Metrics details The PIWI-interacting RNA (piRNA) pathway prevents endogenous genomic parasites from damaging the genetic material of animal gonadal cells are thought to define each species’ piRNA repertoire and therefore its capacity to recognize and silence specific transposon families The unistrand cluster flamenco (flam) is essential in the somatic compartment of the Drosophila ovary to restrict Gypsy-family transposons from infecting the neighbouring germ cells Disruption of flam results in transposon de-repression and sterility yet it remains unknown whether this silencing mechanism is present more widely we systematically characterise 119 Drosophila species and identify five additional flam-like clusters separated by up to 45 million years of evolution Small RNA-sequencing validated these as bona-fide unistrand piRNA clusters expressed in somatic cells of the ovary where they selectively target transposons of the Gypsy family our study provides compelling evidence of a widely conserved transposon silencing mechanism that co-evolved with virus-like Gypsy-family transposons Retrotransposons replicate via RNA intermediates and are further subdivided into non-LTR elements including short interspersed nucleotide elements (SINEs) and long interspersed nucleotide elements (LINEs) which share similarity to endogenous retroviruses (ERVs) LTR transposons and ERVs both encode gag and pol open reading frames (ORFs) with ERVs and specialised retroelements (also known as errantiviruses) such as gypsy and ZAM also possessing an envelope (env) gene The env gene allows virus-like particle formation and cell-to-cell transposition in addition to the “copy-and-paste” mobilisation mechanism intrinsic to all LTR TEs their function and content varies with not all species showing enrichment of transposon remnants a Cartoon of a developing Drosophila egg chamber with an active transposon invasion from the soma (top) Somatic follicle cells lining the egg chamber are shown in green and germ cells are shown in beige Transposon transcripts (purple) originating from somatic cells enter the germ cells (bottom Once reverse transcribed and transported into the nucleus Transposon copy number increases over multiple generations until a transposon is inserted in antisense orientation into flam (step 2 b Cartoon of a Drosophila egg chamber in which transposon invasion is halted (top) A piRNA precursor transcript is produced from the flam locus in somatic cells (bottom The precursor is processed into piRNAs and loaded into Piwi proteins (step 4 The Piwi-piRNA complex enters the nucleus where it recognises transposon transcripts by sequence complementarity and instruments their co-transcriptional repression (step 5 here we systematically searched for flam-like unistrand piRNA clusters within the Drosophila subgenera Sophophora and Drosophila Our results highlight their unique characteristic architecture and specificity in regulating somatically active LTR elements particularly those carrying an envelope protein that facilitates transfer to germ cells our study suggests a conserved and essential role of somatically expressed unistrand piRNA clusters in the suppression of ERVs across the entire Drosophila genus a Cartoon showing synteny analysis pipeline melanogaster (dm6 genome) flam region with transposon annotation by RepeatMasker (RM) displaying some neighbouring genes used for synteny analysis The pie chart to the right indicates LTR content per strand in the cluster region c MCScan plot showing gene and flam synteny between D biarmipes (GCF_018148935 genome) flam region e Phylogenetic tree (left) representation of the melanogaster indicating the respective size of their flam-syntenic loci in kb (right) Source data are available in the source data file In conclusion, the flam locus likely appeared between 13.3 and 15.1 million years ago (MYA), following the emergence of the elegans/rhopaloa subgroups, and was detected in 12 species (Fig. 2e) despite largely conserved gene synteny in the region and the widespread presence of Gypsy-family elements in Drosophilids prompted the question whether analogous flam-like unistrand piRNA clusters exist elsewhere in the genomes of these species a Genome-wide detection of flam-like loci in D ficusphila using a sliding window approach minus strand) and total repeat content (grey) is shown across the whole genome (100 kb bins) and the de novo identified flamlike1 region ficusphila (GCF_018152265 genome) flam-syntenic region (black bar) with transposon annotations by RepeatMasker (RM) and EDTA Uniquely mapping piRNA (cpm) and total RNA levels (ln(cpm+1)) are presented (green/orange c Relative piRNA size distribution of piRNAs mapping sense (light brown) and antisense (dark brown) to D d Ping-pong signature for piRNA pairs mapping onto the flam-syntenic region f Relative piRNA size distribution of sense (light brown) and antisense (dark brown) piRNAs mapping to D g Phasing signature (3’ end to 5’ end distance) for piRNAs mapping onto flamlike1 h Zoom-in on genic region indicating presence of a piRNA cluster in the flam-syntenic region in D i Macrosynteny plot indicating gene synteny between D ficusphila highlighting flam (red) and flamlike1 (blue) j Zoom-in on genic region indicating the absence of a piRNA cluster in flamlike1-syntenic region in D As D. ficusphila appears to possess a dual-strand cluster in place of the flam locus, it either lacks somatically expressed LTR transposons or controls these TEs by other means. The presence of Gypsy family elements in all investigated genomes strongly indicates that D. ficusphila has somatically expressed transposons (Fig. S3c) We therefore set out to identify non-syntenic unistrand piRNA clusters in D ficusphila that resemble flam in terms of its size Gypsy-family TE content and strong enrichment for transposon insertions to be oriented on one genomic strand oshimai flamlike2 region with transposon annotation by EDTA (blue e Phylogenetic tree representation highlighting flamlike1 flamlike2 and flamlike4 presence and flamlike3 and flamlike5 conservation across Drosophila species pseudoobscura flamlike5 syntenic dual-strand cluster consistent with expression in somatic cells that lack the machinery needed to express and export transcripts from dual-strand clusters since sRNA-seq from whole ovaries captures a mixture of both somatic and germline piRNAs it remained uncertain if these unistrand piRNA clusters actually operate in the soma g Phylogenetic tree summarising all studied somatic piRNA clusters across all analysed species (n = 119) Cluster size represents the mean across all assemblies Species names in bold have sRNA-seq data to validate their expression Interestingly, we found that D. yakuba and D. erecta deviated from this pattern, displaying somatic expression at the 5’ end and germline expression towards the 3’ end of the flam-syntenic region (Fig. S14b, c) this indicates that all identified flam-like loci produce antisense piRNAs capable of targeting transposons and that they are expressed primarily in the somatic follicle cells of the ovary this indicates a strong selective pressure to maintain production of transposon-targeting piRNAs in somatic follicle cells a Boxplot showing fraction of interspersed repeat content for the indicated repeat classes Each data point represents one species (n = 119) Species with multiple genome assemblies are represented by their mean b Boxplot showing the number of subfamilies detected per LTR family with either gag + pol (left) or gag + pol + env (right) ORFs Each data point corresponds to one species (n = 119) sense) to total transposon content across all annotated flam-like clusters Gypsy elements are shown in red (antisense) or blue (sense) and other LTR elements are shown in grey Clusters are grouped by synteny as indicated to the right Species and genome assembly (alphabetically sorted) are indicated to the left but showing LTR content across flam and major dual-strand clusters in D Cluster strand was defined according to total transposon content (light grey) e Boxplot showing strand bias defined as sense strand minus antisense strand contribution to total transposon content for transposons classified as LTR Strand bias is shown across all annotated flam-like clusters (left n = 48) or major dual-strand clusters in D melanogaster and proTRAC de novo predicted clusters (right The means were compared using a two-sided Student’s t Test f Boxplot displaying Gypsy versus other LTR coverage against the genomic average across different unistrand clusters Each point corresponds to one cluster in one genome assembly g Scatterplot showing Gypsy enrichment against env enrichment in unistrand clusters from the indicated species (see “Cluster content analyses” in the Methods for details) Only high-quality LTR transposons are included in the analysis (both gag and pol and at least one good genomic hit) suggesting that similarities in piRNA populations between species is indicative of shared transposon burden To further our understanding of how transposons are regulated by flam-like clusters we characterised the individual transposons that are controlled by each cluster These species were selected based on the availability of both whole ovary and soma-enriched sRNA-seq and RNA-seq As controls we used the dual-strand 42AB in D Genomic origin of piRNAs that are antisense to transposons in D Barplots (right) display the number of uniquely mappable piRNAs against each TE in soma-enriched (a The TEs are arranged in decreasing order following their somatic-to-germline enrichment The number of piRNAs that map to the indicated clusters are coloured according to cluster strand (sense red) and piRNAs mapping elsewhere in the genome are shown in grey Labels are shown for subfamilies that are exclusively controlled (>90% of piRNAs) by a cluster and best hit to known TEs are indicated if available (80/80/80 rule) Boxplots (left) summarise the fraction of piRNAs antisense to individual transposons derived from each cluster Total cluster-derived piRNA abundance (white) are further subdivided into the sense (blue) and antisense (red) cluster strands The number of transposon subfamilies covered by each cluster (>10 reads) are indicated under each boxplot Pooled counts from 2–4 biological replicates Our identification of flam within the suzukii subgroup together with the absence of any piRNA cluster at the flam-syntenic region in and beyond the rhopaloa subgroup places the emergence of flam between 13.3 and 15.1 MYA long before the melanogaster subgroup separated from the remainder of the melanogaster group around 6.8 MYA We speculate that all Drosophila species use flam-like piRNA clusters in a somatic branch of the pathway that specifically evolved to repress ERVs While flam-like clusters have not been detected in all species we have consistently identified somatic unistrand piRNA clusters in all species where we performed soma-enriched sRNA-seq we did not observe any Drosophila species lacking the presence of env-containing Gypsy-family elements More unistrand piRNA clusters are therefore likely to be discovered as we gain access to more sequencing data and improved genome assemblies in the future Most flam-like unistrand clusters reported here follow the pattern of a single large locus We hypothesise that in addition to being the most efficient way of stopping TE invasion as disruption of flam-like piRNA clusters likely result in sterility The recurring presence of unistrand clusters across the Drosophila genus strongly argues for an essential role of these loci perhaps as a means to produce piRNAs in the soma without access to the germline piRNA expression and export machinery Conversion between unistrand (a) and dual-strand (b) piRNA clusters Transposons are present either in sense (blue) or antisense (red) orientation relative to the cluster transcript(s) Produced piRNAs mapping to the sense (green) or antisense (orange) strand are shown Once a promoter active in the soma is gained (a) selection will favour antisense insertions to ensure that transposon-complementary piRNAs are produced the cluster can only be transcribed in germ cells where the germline-specific branch of the piRNA pathway produces transcripts from both strands The strand bias is therefore lost over evolutionary time d Selective constraints acting on unistrand piRNA clusters Transposon insertions in sense orientation are tolerated towards the 3ʹ end (c) but are rarely observed at the 5ʹ end (d) This may indicate that the region closer to the promoter is under stronger selective pressure insertions in sense orientation may introduce polyadenylation signals causing early transcription termination abolishing the production of essential piRNAs targeting specific TEs (d) Although their transcriptional regulation may differ the recurrent emergence of flam-like loci across the Drosophila genus and the wider presence of unistrand clusters within in the animal kingdom hints at convergent evolution where this mechanism is best equipped to antagonise TE mobilisation our study opens the door to understanding the co-evolution between virus-like Gypsy-family transposons and the host defence mechanisms that silence them Further characterisation of these novel piRNA clusters as well as the piRNA pathway machinery in these species will allow us and others to test several long-standing hypotheses regarding piRNA cluster emergence and the licensing of their transcripts for piRNA biogenesis Assemblies for 36 species annotated by the NCBI Eukaryotic Genome Annotation Pipeline (listed on https://www.ncbi.nlm.nih.gov/genome/annotation_euk/all) from any species within the Drosophila genus were downloaded on three separate occasions (2020-10-17 Only the most recently annotated genome assembly is listed for each species and as a result 19 species were represented by a single assembly and 15 species were represented by two different assemblies Since the clusters themselves are not conserved, we used a synteny analysis (see “Synteny_clusters” at https://github.com/susbo/Drosophila_unistrand_clusters) melanogaster genome as a reference and extracted the 20 unique up- and downstream genes we extracted the coding sequence (protein-coding genes) or the full transcript (all others) we mapped these sequences onto the genome of interest using blat (v36x6 -minIdentity=25) and filtered the results to keep the best hit (pslCDnaFilter we constructed a candidate list with all genomic regions that had at least two gene hits within 1 Mb These candidate regions were then manually inspected for the presence of a transposon-rich area at the expected syntenic location Clusters running into assembly breakpoints were labelled as either 5’ or 3’ depending on whether they were located next to up- or downstream genes De novo identified clusters (proTRAC) were converted from GTF to GenePred RNA-seq and sRNA-seq tracks were displayed as standard bigWig tracks produced by deepTools All genome browser shots shown in this study were made by exporting the assembly hub display as a pdf followed by manual refinement to enhance readability Mappability tracks for the sRNA-seq were constructed by generating all possible 26-mers from each genome (bedtools -S -n 2 -M 1 --best --strata --nomaqround --chunkmbs 1024 --no-unal) and converting the alignments to bigWig using deepTools bamCoverage (v3.3.2 --binSize 1 --normalizeUsing None --scaleFactor 0.038461) This will construct a per-nucleotide signal between 0 and 1 representing the ability to uniquely map reads to each position An initial de-novo transposon library was built using EDTA (v1.9.3, --sensitive 1 --anno 1 --evaluate 1)37 with transposons of all types being detected some runs failed when one of the types were missing and we manually resumed EDTA at the next type for these genomes and Zind-d101g_BS02) that failed to run with EDTA v1.9.3 did run successfully with v1.9.6 Dwas-d101g) that had problems with v1.9.3 still had to be resumed with v1.9.6 due to not detecting any LTR transposons To search for flam-like clusters, we developed a search strategy based on the known enrichment of LTR transposons arranged in the same orientation in flam (see “De-novo_clusters” at https://github.com/susbo/Drosophila_unistrand_clusters) repeat annotations from the EDTA were used The repeats were separated based on strand retaining either only LTR transposons or all transposons with a predicted class (i.e. Overlapping annotations were combined (bedtools merge) and strand-specific transposon coverage was computed (bedtools coverage) across the genome using a 100 kb sliding window with a 5 kb step size (bedtools makewindows Each genome was manually inspected for regions enriched in LTR transposons and located outside of centromeric or telomeric regions This analysis was strongly contingent on assembly quality but we nevertheless identified 15 clusters that fulfilled the outlined criteria including several corresponding to flam across the D Six of the initial candidates were found outside of the D two species had publicly available sRNA-seq data and both produced large amounts of piRNAs from one strand only We therefore concluded that the approach was working Synteny analysis using these five clusters as starting points (described below) revealed that D All Drosophila species were maintained at room temperature. The origin of each species and their food requirements are indicated in Supplementary Data 8 Small RNAs were isolated from 16 species (2–3 replicates each) using the TraPR Small RNA Isolation Kit (Lexogen; catalogue nr 128.24) following the manufacturer’s instructions sRNA libraries were generated using the Small RNA-Seq Library Prep Kit (Lexogen; catalogue nr Both primers A3 and A5 as well as the primer RTP were used at 0.5x Library size distribution was analysed on an Agilent TapeStation system using a High Sensitivity D1000 ScreenTape (Agilent Technologies; catalogue nr 5067-5584) with High Sensitivity D1000 Reagents (Agilent Technologies; catalogue nr Libraries were pooled in equal molar ratio quantified with KAPA Library Quantification Kit for Illumina (Kapa Biosystems; catalogue nr KK4873) and were sequenced 50 nt paired-end on an Illumina NovaSeq 6000 or 75 nt single-end on an Illumina MiSeq sequencing platform generating 33 (±20) million reads per library 75-100 ovary pairs were dissected in ice-cold PBS Ovaries were dissociated for 18 min in 0.25% Trypsin (Sigma-Aldrich; catalogue nr Dissociated tissue was pushed through a 40 µm nylon mesh (Greiner Bio-One; catalogue nr 542040) washed with equal volume Schneider 2 medium (Thermo Fisher Scientific; catalogue nr Pelleted cells were directly used as input for sRNA isolation using the TraPR Small RNA Isolation Kit (Lexogen; catalogue nr KK4873) and were sequenced 50 nt paired-end on an Illumina NovaSeq 6000 sequencing platform generating 43 (±25) million reads per library The BAM files were converted to bigWig using bamCoverage from deepTools71 (v3.3.2 --binSize 1 --ignoreForNormalization chrM --normalizeUsing CPM --exactScaling --skipNonCoveredRegions --minFragmentLength 23 --maxFragmentLength 30) and additionally ‘--filterRNAstrand’ to separate the two strands ‘--scaleFactor’ to scale counts per million to reflect all mapped reads and optionally ‘--minMappingQuality 50’ when extracting uniquely mapped reads RNA-seq libraries were produced using NEBNext Ultra Directional Library Prep Kit for Illumina (New England BioLabs; catalogue nr following the manufacturer’s instructions for rRNA depleted RNA KK4873) and sequenced paired-end 50 nt on an Illumina NovaSeq 6000 generating 25 (±11) million reads per library The soma-enrichment RNA-seq libraries were generated for 5 species (2 replicates each) Enrichment for somatic cells was done identically as described for the soma-enriched sRNA-seq libraries except that 35-50 ovary pairs were used as starting material Pelleted cells were directly used as input for RNA isolation using the TRIzol (Thermo Fisher Scientific; catalogue nr RNA was treated with DNase (New England BioLabs; catalogue nr M0303) followed by ribosomal RNA depletion using RiboPOOL (siTOOLs Biotech; catalogue nr dp-K024-000007) following the manufacturer’s protocol KK4873) and sequenced paired-end 50 nt on an Illumina NovaSeq 6000 generating 42 (±7.1) million reads per library we calculated the ping-pong signature using a 5’ end overlap score for overlap x nt as where ni is the number of 5’ ends mapping at the plus strand position i and mi+x is the number of 5’ ends mapping at the minus strand position i + x The fraction of overlapping reads involved in ping-pong was calculated as s10/(s1 + …+s20) A z10 score was defined as (s10-mean(s1,…,s9,s11,…,s20))/stdev(s1,…,s9,s11,…,s20) we calculated a 3ʹ to 5ʹ end score for distance y as where ni is the number of 3’ ends mapping at position i and mi+y is the number of 5’ ends mapping at position i + y at the same strand The fraction of closely mapped reads with phasing signature was calculated as h1/(h1 + …+h20) A z1 score was calculated as (h1-mean(h2,…,h20))/stdev(h2,…,h20) Phasing calculations were done for the plus and minus strand separately Synteny analysis for flam-like clusters was performed using the same strategy as for flam except that Augustus gene predictions (v3.3.2 --species=fly --UTR=off --singlestrand=true) were used instead of FlyBase annotations The MAKER-masked genome from the EDTA output was used as genome input to Augustus Full transcript and coding sequences were extracted from the annotations Sequences with strong hits to the raw transposon libraries were excluded (blat -q=dna -t=dna -minIdentity=25; pslCDnaFilter -minCover=0.2 -globalNearBest=0) and gene predictions shorter than 200 nt were excluded The blat identity threshold was reduced to 20 the closest flanking genes displayed good conservation and we used these to search for syntenic regions using our UCSC Genome Browser session ATAC-seq was performed for nine species similar as described in77 6-12 ovary pairs of yeast-fed flies were dissected in ice-cold PBS and centrifuged for 5 min at 500 g at 4 °C Ovaries were lysed in Resuspension Buffer (RSB 3 mM MgCl2 in nuclease free water) containing 0.1% NP40 and 0.01% Digitonin and washed out with cold RSB containing 0.1% Tween-20 The transposition reaction was performed with 0.33x PBS 1x TD buffer and 100 nM transposase (Illumina Tagment DNA Enzyme and Buffer Small Kit; catalogue nr Samples were incubated for 1 h at 37 °C in a thermomixer mixing at 1000 rpm The transposed fragments were isolated using the DNA Clean and Concentrator-5 Kit (Zymo Research; catalogue nr Library was PCR amplified for 5 cycles using the NEBNext High-Fidelity MasterMix (New England BioLabs; catalogue nr M0541S) followed by qPCR amplification to determine the exact number of additional cycles required for optimal library amplification Amplified DNA library was purified using the DNA Clean and Concentrator-5 Kit (Zymo Research; catalogue nr D4014) and further cleaned using AMPure XP beads (Beckman Coulter; catalogue nr 100-600 bp fragments were selected on a 2% agarose gel cassette using the Blue Pippin (Sage Science; catalogue nr Library size distribution was analysed on an Agilent 2100 bioanalyzer using the High Sensitivity DNA Kit (Agilent Technologies; catalogue nr KK4873) and were sequenced 50 nt paired-end on an Illumina NovaSeq 6000 platform or Illumina MiSeq sequencing platform generating 4.5-25.6 million paired-end reads per library with ‘-repeatmasker’ set to RepeatMasker annotations generated by EDTA and with ‘-geneset’ set to NCBI gene predictions Clusters within 40 kb from each other were combined for the analyses melanogaster clusters were mapped onto each genome using BLAT (v36x6 filtered to retain only the best hit (pslCDnaFilter -minCover=0.2 -globalNearBest=0.0) and the predicted clusters were subsequently annotated by how many of these genes that were within 1 Mb Cluster predictions used in the soma-enrichment analysis were performed using the same strategy but restricted to libraries generated for this study and using either only soma-enriched or only total sRNA-seq libraries (2–3 replicates per species and library type) Clusters identified using either somatic or total libraries were concatenated and any clusters within 40 kb from each other were merged Clusters of size <35 kb were discarded to enable analysis of strand biases across major clusters Total piRNA coverage per cluster was normalised to counts per million and calculated for soma-enriched and total libraries separately Somatic clusters were defined as clusters with at least 2-fold soma enrichment over total libraries and germline clusters were defined as being higher expressed in the total ovary libraries In addition to the consensus sequences obtained from EDTA we also used the RepeatModeler (v2.0.1) output within the EDTA folders to improve detection of LINE elements We reasoned that we could not provide a list of known LINEs to EDTA since that would mainly reflect melanogaster transposons and would bias the comparisons between the melanogaster subgroup and other species EDTA and RepeatModeler consensus sequences were combined and further processed using a custom pipeline (see “Transposon_libraries” at https://github.com/susbo/Drosophila_unistrand_clusters) and remaining sequences were clustered using cd-hit-est (v4.8.1 -G 0 -g 1 -c 0.90 -aS 0.90 -n 8 -d 0 -b 500) to combine any sequences with ≥90% identity across ≥90% of the length Custom scripts were used to select one representative sequence from each cluster maximising both the number of high-quality genomic hits (blastn filtered to cover at least 50% of the query sequence) and the length of the sequence Sequences with fewer than 2 high-quality genomic hits were removed from the transposon libraries To prioritise sequences and to detect known transposon domains gag and pol ORFs from RepeatPeps.lib in RepeatMasker (v4.1.2) using blastx (v2.10.0 Sequences covering at least 50% of the full peptide domain were considered true hits which were considered to belong to the same subfamily This was repeated using 90/80/90 and 80/80/80 thresholds to detect more distant similarities The curated and annotated transposon consensus sequences have been made available (https://github.com/susbo/Drosophila_TE_libraries) --max-seeds 100 -q -k 1 -p 10 --no-unal --new-summary --summary-file) the median alignment rate was subtracted from each library to reduce false hits driven by abundant non-coding RNAs To determine whether clusters were more likely to have captured Gypsy-family LTR transposons compared with other LTR transposons we defined a Gypsy enrichment ratio as Gypsy is the set of all Gypsy-family transposon subfamilies Captured is the set of all transposon subfamilies found inside the cluster region and Not captured is the set of all other transposon subfamilies Reads mapping uniquely to each genome were intersected with cluster coordinates using bedtools intersect Resulting counts were normalised to the total number of reads mapping to each genome For the cluster content analysis (Fig. 7) we considered only reads of length 24-28 nt that mapped uniquely to the curated transposon libraries a set of 100 piRNA-regulated transposons were defined for whole ovary and soma-enriched ovary by ranking the sequences in the curated transposon library by the number of piRNAs mapping to them across all replicates The rankings were highly similar between whole ovary and soma-enriched ovary and in total 116 to 128 transposon subfamilies were selected per species To enable comparison of the counts in soma-enriched and whole ovary libraries we derived cpm values by normalising the counts to the total number of reads mapping to the genome Soma-enrichment per transposon subfamily was calculated as the difference in cpm between the pooled soma-enriched and whole ovary libraries we further restricted the analysis to reads that also mapped uniquely to the genome assembly and used the read identifiers to assigned transposon identity and transposon strand to each genome-mapping read the reads were intersected to piRNA cluster coordinates (bedtools intersect v2.26.0) in strand-specific mode to allow determination of whether the reads originated from the sense or antisense strand of a cluster dual-strand clusters were assumed to be located on the + strand Metadata for the curated transposon libraries were obtained as described previously under “Construction of curated de novo transposon libraries” To avoid false hits to conserved protein domains and to increase sensitivity compared with sequence-based searches, we employed a synteny-based search strategy (see “Synteny_biogenesis_genes” at https://github.com/susbo/Drosophila_unistrand_clusters) melanogaster genome as a reference and extracted the 20 closest genes up- and downstream genes -maxIntron=500000 -minMatch=2 -minScore=30 -oneOff=1 -minIdentity=10) and filtered the results to keep the best hit (pslCDnaFilter we constructed a hit list with all genomic regions that had at least two hits within 1 Mb from another which was manually inspected for the presence of the gene of interest Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article The flamenco Locus Controls the gypsy and ZAM Retroviruses and Is Required for Drosophila Oogenesis piRNA-mediated regulation of transposon alternative splicing in the soma and germ line Genome surveillance by HUSH-mediated silencing of intronless mobile elements The hush complex cooperates with trim28 to repress young retrotransposons and new genes The role of KRAB-ZFPs in transposable element repression and mammalian evolution PIWI-interacting RNAs: small RNAs with big functions piRNA-guided genome defense: From biogenesis to silencing Discrete small RNA-generating loci as master regulators of transposon activity in drosophila Evolutionarily conserved pachytene piRNA loci are highly divergent among modern humans An ancient transcription factor initiates the burst of piRNA production during early meiosis in mouse testes A single unidirectional piRNA cluster similar to the flamenco locus is the major source of EVE-derived transcription and small RNAs in Aedes aegypti mosquitoes A role for Piwi and piRNAs in germ cell maintenance and transposon silencing in Zebrafish Adaptive evolution leads to cross-species incompatibility in the piRNA transposon silencing machinery The rhino-deadlock-cutoff complex licenses noncanonical transcription of dual-strand piRNA clusters in Drosophila Evolutionary dynamics of piRNA clusters in Drosophila Transcriptional properties and splicing of the flamenco piRNA cluster Export of piRNA precursors by EJC triggers assembly of cytoplasmic Yb-body in Drosophila a gene controlling the gypsy retrovirus of Drosophila melanogaster Gypsy transposition correlates with the production of a retroviral envelope-like protein under the tissue-specific control of the Drosophila flamenco gene The beta heterochromatic sequences flanking the I elements are themselves defective transposable elements Drosophila germline invasion by the endogenous retrovirus gypsy: Involvement of the viral env gene Infection of the germ line by retroviral particles produced in the follicle cells: a possible mechanism for the mobilization of the gypsy retroelement of Drosophila and diversity of retrotransposons at the flamenco locus reflect the regulatory properties of piRNA clusters Recurrent insertion and duplication generate networks of transposable element sequences in the Drosophila melanogaster genome piRNA clusters need a minimum size to control transposable element invasions Functional adaptations of endogenous retroviruses to the drosophila host underlie their evolutionary diversification Conserved piRNA expression from a distinct set of piRNA cluster loci in eutherian mammals Specialized piRNA pathways act in germline and somatic tissues of the Drosophila ovary Highly contiguous assemblies of 101 drosophilid genomes Highly contiguous genome assemblies of 15 drosophila species generated using nanopore sequencing History of the discovery of a master locus producing piRNAs: the flamenco/COM locus in Drosophila melanogaster A heterochromatin-dependent transcription machinery drives piRNA expression Channel nuclear pore complex subunits are required for transposon silencing in Drosophila Benchmarking transposable element annotation methods for creation of a streamlined A slicer-mediated mechanism for repeat-associated siRNA 5′ end formation in Drosophila The absence of core piRNA biogenesis factors does not impact efficient transposon silencing in Drosophila proTRAC-a software for probabilistic piRNA cluster detection Poised for contagion: evolutionary origins of the infectious abilities of invertebrate retroviruses Evolution and phylogeny of insect endogenous retroviruses Gypsy endogenous retrovirus maintains potential infectivity in several species of Drosophilids Trapping a somatic endogenous retrovirus into a germline piRNA cluster immunizes the germline against further invasion Reactivation of a somatic errantivirus and germline invasion in Drosophila ovaries on the neo-Y chromosome of Drosophila miranda Requirements for multivalent Yb body assembly in transposon silencing in Drosophila a major site for Piwi-associated RNA biogenesis and a gateway for Piwi expression and transport to the nucleus in somatic cells Dynamics of Transposable Element Invasions with piRNA Clusters Large Drosophila germline piRNA clusters are evolutionarily labile and dispensable for transposon regulation Natural variation of piRNA expression affects immunity to transposable elements Rapid evolution of piRNA clusters in the Drosophila melanogaster ovary The Drosophila HP1 homolog Rhino is required for transposon silencing and piRNA production by dual-strand clusters The Cutoff protein regulates piRNA cluster expression and piRNA production in the Drosophila germline A transcriptome-wide RNAi screen in the Drosophila ovary reveals factors of the germline piRNA pathway Daedalus and gasz recruit armitage to mitochondria bringing piRNA precursors to the biogenesis machinery A heterochromatin-specific RNA export pathway facilitates piRNA production Rapid evolutionary diversification of the flamenco locus across simulans clade Drosophila species Widespread introgression across a phylogeny of 155 Drosophila genomes a database of repetitive elements in eukaryotic genomes Adaptive seeds tame genomic sequence comparison DrosoPhyla: Resources for drosophilid phylogeny and systematics Maternally deposited germline piRNAs silence the tirant retrotransposon in somatic cells Rapid evolutionary dynamics of an expanding family of meiotic drive factors and their hpRNA suppressors Deep experimental profiling of microRNA diversity Pan-arthropod analysis reveals somatic piRNAs as an ancestral defence against transposable elements Run or Die in the Evolution of New MicroRNAs-Testing the Red Queen Hypothesis on De Novo New Genes piRNA silencing contributes to interspecies hybrid sterility and reproductive isolation in Drosophila melanogaster miRBase: from microRNA sequences to function deepTools2: a next generation web server for deep-sequencing data analysis Specialization of the Drosophila nuclear export family protein Nxf3 for piRNA precursor export melanogaster modENCODE transcriptome annotation The genome of drosophila innubila reveals lineage-specific patterns of selection in immune genes Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype PiRNA-guided transposon cleavage initiates Zucchini-dependent An improved ATAC-seq protocol reduces background and enables interrogation of frozen tissues Fast and accurate short read alignment with Burrows-Wheeler transform and validation of bias in ATAC-Seq data with ataqv BEDTools: a flexible suite of utilities for comparing genomic features Considering transposable element diversification in de novo annotation approaches A unified classification system for eukaryotic transposable elements van Lopik, J. et al. Unistrand piRNA clusters are an evolutionarily conserved mechanism to suppress endogenous retroviruses across the Drosophila genus, Drosophila_TE_libraries, Zenodo, https://doi.org/10.5281/zenodo.8377332 (2023) van Lopik, J. et al. Unistrand piRNA clusters are an evolutionarily conserved mechanism to suppress endogenous retroviruses across the Drosophila genus, Drosophila_unistrand_clusters, Zenodo, https://doi.org/10.5281/zenodo.8377341 (2023) Download references We thank Hannon group members for fruitful discussions Holly Goodrick for help with library preparation and Emma Kneuss for providing early access to unpublished sRNA-seq libraries We thank the Scientific Computing core at the CRUK Cambridge Institute for HPC resources and the Genomics core for sequencing services GJH is a Royal Society Wolfson Research Professor (RSRP\R\200001) by Cancer Research UK (G101107) and the Wellcome Trust (110161/Z/15/Z) The following species were kindly provided by the indicated laboratories: D virilis from Simon Collier at the Department of Genetics These authors contributed equally: Jasper van Lopik Susanne Bornelöv & Benjamin Czech Nicholson designed the experiments and interpreted the results performed ATAC-seq experiments and analysis performed all other wet-lab work and preliminary computational analysis except the MCScan analysis that was performed by MAT All authors read and approved the final version The authors declare no competing interests reviewer(s) for their contribution to the peer review of this work Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41467-023-42787-1 Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Sign up for the Nature Briefing newsletter — what matters in science — Several roads in Robinson Township were impassable Sunday as a significant ice jam that formed on the Grand River sent the river over its banks The ice jam at the river near Van Lopik Avenue and Limberlost Lane has since shifted west of the M-231 bridge and as of Monday morning was near Connor Bayou Much of the floodwater east of the M-231 bridge receded back into the river when the ice jam moved As of Monday morning no homes in Robinson Township had been damaged by the flooding but officials in Ottawa County are closely watching the ice jam for potential flash flooding in the next several days The National Weather Service issued a flood warning for the area for Monday morning through Tuesday evening warning that low-lying areas near Connor Bayou could flood suddenly and rapidly "All bets are off with an ice jam," said Ottawa County Emergency Management Director Lou Hunt describing the phenomenon's unpredictability More: Officials warn of possible ice jams on Grand River after Wednesday warm-up, rain Flooding can occur upstream from the ice jam when it blocks river flow The ice jam can also cause flash flooding downstream if there is a sudden release of the ice and water rushes through The Michigan State Police were to conduct an fly-over of the Grand River on Monday to survey the river Hunt said the county is most concerned about flooding impacts in Robinson Township where there are low-lying homes that could be affected if a flash flood occurs They will be closely tracking the ice jam as it is expected to move west down the river over the next several days "The rises in the water levels are pretty unpredictable," Hunt said "At Van Lopik and Limberlost they didn't come up tremendously high but they were fast They came up quickly and they came down quickly so we really want people to keep their eyes on it and make a plan to quickly move to higher ground." Hunt also expected the river to swell through the day Monday as the melted snow and rain from last week's warmer weather throughout the Grand River watershed makes its way to the Ottawa County section of the river The National Weather Service warned drivers not to attempt to drive through flooded roads in the area Ottawa County EMA advised residents that live in flood-prone areas near the ice jam to pack essentials in a "go bag" to be ready to evacuate quickly — Contact reporter Carolyn Muyskens at cmuyskens@hollandsentinel.com and follow her on Twitter at @cjmuyskens.  .st1{fill-rule:evenodd;clip-rule:evenodd;fill:#2a2a2a}By Terry DeBoer | grentertainment@mlive.comJessica Scott | The Grand Rapids PressJessica Vogel preforms as Anne Shirley grabbing at Brenda VanderArk's character Rachel Lynde while Mary Ann Heinen plays Marilla Cuthbert during a scene from Master Arts Theatre performance of the musical "Anne of Green Gables."BYRON TOWNSHIP -- Master Arts Theatre had to do some scrambling to stage its fall musical "We had been planning on doing 'The Music Man,' but we learned that a national (theater) company would be touring that show," said director Kathy Van Lopik That meant the local community theater group could not secure the rights necessary to perform the legendary musical "I was aware of the 'Anne of Green Gables' show and thought it fit our theater and clientele well so we decided to go ahead with that one," said the director Van Lopik did a reshuffling of the actor roster She also made a few phone calls and came up with the final 29-member cast for "Anne of Green Gables." The lead role of Anne Shirley -- the teenage orphan who comes to Green Gables farm in Prince Edward Island The Calvin Christian High School student had been selected for a lesser part in "Music Man." Admission: Reserved seat tickets $17, $15 for students and senior citizens. Some dates may already be sold out. Call 455-1001 or visit masterarts.org "It's been really cool seeing the show come together," Vogel said of rehearsals that began in late July "I have to make myself remember to let go -- to throw every emotion I can into it," she said of her preparations for the role of Anne an imaginative teen who became the subject of gossip and jealousy in a small Canadian farm community of a century ago Van Lopik has seen the musical adaptation that is staged each year in Prince Edward Island But she could not find a theater in Michigan that has presented the tale's musical version "The story is full of both comical and tender moments and the music helps that along very well," Van Lopik said of the account of how Anne wins over her adopted family and surrounding residents even in the face of difficult choices Master Arts' smaller theater prevented construction of a two-story farm house set used in the original production The director had to rethink how to stage the play using a split set -- the large home's main floor is on one side of the stage "You can put music beds under some of the scene transitions," Van Lopik said of a tactic often used "But we've found a way to spot(light) one room and then another allowing for the time the character would need to move through a large house." is a veteran of Master Arts drama workshops and stage productions She had the lead in last fall's "I Remember Mama." She has been in all but one of her high school stage shows -- one that conflicted with her role in a concurrent Master Arts production "There are a lot of kids in this show -- I was surprised at how many," she said "But we've really bonded as a cast Use of and/or registration on any portion of this site constitutes acceptance of our User Agreement, (updated 8/1/2024) and acknowledgement of our Privacy Policy, and Your Privacy Choices and Rights (updated 1/1/2025) © 2025 Advance Local Media LLC. All rights reserved (About Us) The material on this site may not be reproduced except with the prior written permission of Advance Local Community Rules apply to all content you upload or otherwise submit to this site YouTube's privacy policy is available here and YouTube's terms of service is available here Ad Choices .st1{fill-rule:evenodd;clip-rule:evenodd;fill:#2a2a2a}By Community News | The Muskegon Chroniclephoto/Tri-Cities Historical MuseumA popular business in the early 1900s was Garrett Ekkens' summer store on Lake Avenue Ekkens' had a main store in downtown Grand Haven but he opened a Highland Park outlet to cater to summer tourists.For over 100 years generation upon generation has been returning to their tree-topped cottages nestled in the dunes known as Highland Park with a mix of seasonal cottages and year-round homes the special charm and history still remain Stretching from Lake Avenue on the north to Grand Avenue on the south shaded boardwalks and paths wind around the dunes to recall memories when residents had to hand carry groceries and suitcases to their cottage because it could not be reached by automobile Many families are continuing their legacy in cottages that their parents or grandparents owned Highland Park has been described as a “singular neighborhood within the city even perhaps unique within the state of Michigan.” Through the years the residents have fought to preserve its unique character by not altering the natural dune environment Residents have been celebrating the 125th anniversary with several social gatherings culminating with an 'end of the summer' breakfast in September Below is a history of the individual Highland Park cottages compiled by local historian Wally Ewing: and Grand Haven proved that the area could attract large numbers of vacationers Grand Haven and other cities along the Lake Michigan shore were becoming frequented by well-to-do summer visitors from large urban centers such as Detroit and many of them enjoyed staying in Grand Haven for the entire season The City of Grand Haven for $350 had acquired title to a large tract of land 1886 the City leased a portion of this land for a thirty-year term to the newly formed Highland Park Association The Association raised $5,000 capital by selling shares at $25 Most of the charter members bought one or two shares The Association made plans for a resort community of cottages overlooking Lake Michigan Resorters arrived by boat from Chicago or by the interurban from Grand Rapids and other Michigan cities Teams of horses and the backs of laborers like John Vyn were used to carry trunks and other baggage from the depots to the cottages With primitive roads and limited means of transportation workers had to carry lumber up the hills by hand exposing residents to chilling drafts once the warm breezes of August had departed for another year Because so many of the cottages were built on the sides of steep dunes and in ravines long wooden stairways and boardwalks were constructed But life during the summer months was grand The Highland Park Tennis Club's grass courts attracted some of the world's best players Gerrit Ekkens drove up the road from town with a wagon full of provisions from his grocery always sure to have an ample supply of his famous cheese residents could pick up groceries for themselves and tour the downtown area at the same time The mail service experienced a problem unique to the Highland Park area For a long time there were no house numbers and the cottages were simply identified by name Some charming examples included Highland Castle and some of the nonresident buildings have been removed Numerous other changes have been made to the Highland Park area but forever present will be the pleasant memories of summers spent by the shore These are some Highland Park Memories that have been recorded: "Highland Park was such a friendly sort of 'big family' association as I grew up August Boseker ran the ice cream parlor and candy stand in such a pleasant way and each Sunday afternoon there would be a dance pavilion After the concert there was always a mad rush for the ice cream parlor." "A trip to the cottage was not complete without a trip to the woods through Fairyland an enchanted open sand bowl in the middle of the woods where fairies and brownies lived he loved kids and would give us raw sugar or mangoes or some other tropical treat He kept rattlesnakes in formaldehyde in a bottle." there used to be a well where we all went to get our drinking water I think it was simply pumped up from the lake without going through any sort of treatment." went down the beach to a place north of where the state park is now located to whistle for the Coast Guard boat to pick them up and take them across the channel to the Coast Guard Station for swimming lessons taught by the Coast Guard men by tying a rope around them and dangling them in the channel." it was registered by the State of Michigan an historic site Four years earlier a preservation consultant had said "The contrast of the density of development on the dune ridges with the pristine forest of the slopes and valleys creates an intimate environment which is at once a recreational and educational resource." One of the most popular destinations for big city tourists was the elegant Highland Park Hotel where some families made return visits for three generations Built between 1888 and 1891 [one source says it opened on July 4 the two-story frame hotel featured 36 rooms in the main lodge and 15 more in the annex Porches on both levels extended the full length of the building From 1899 [1894] to 1908 [1914] Martha and George McBride managed the resort [Another report claimed that Chicago residents Sarah Sweetland and her two sons purchased the hotel in 1906 and it was these people who built the Annex.] In 1893 the rates were $2.00 per day with "special terms by the week given to families desiring to stay during the season." The advertisement also boasted that the resort offered "GOOD FISHING BOATING AND BATHING." At some point a portion of the long porch was converted to rooms and running water replaced the commode with pitcher and bowl which then rented for $5 a night during the season beginning in late July and ending in early September and the hotel was owned and managed by Edward M 1947 the Langrells sold the hotel to Mell Wright who owned Wright's Grill on Washington Street The hotel was destroyed by a fire on the night of December 20 but the annex survived as a bed and breakfast In 1888 Martha and George McBride of Grand Haven built this cottage   McBride was one of the original shareholders and directors of the Highland Park Association The McBrides managed the Highland Park Hotel from 1899 to 1908 Hazen and William Blakeslee purchased Cozy Corner for $2,500 Later improvements included bringing in a refrigerator to replace the icebox and a gas range replaced the old metered gas stove that would ignite when a quarter was inserted A bathroom was added shortly after World War II and a few years later an upper level was added This large eight-bedroom cottage was bought by Maribeth Van Vliet This stately cottage was owned by the Stobie/Mare family from St Louis for nine decades Helen Mare Dean's grandmother's father and brother built it in 1896 Yvonne Lynch and John McKewan bought the cottage in 1995 and gave it the name Trillium Bluff The cottage at this address was built about 1912 Fifteen years later the Strand family bought it Owners in 2000 were Charles and Barbara Soderquist In the early 1900s this cottage was owned by an actress who had the lead role in a play on Broadway called Seven Keys to Baldpate who moved to Grand Haven to take a job around 1917 as Chief Clerk with the Grand Trunk Railway for the next 12 years The actress and Edwin sold the place to Norma's father The cottage later was owned by Charles Everest who served in that capacity from 1936 to 1953 During a raging thunderstorm around 1980 the structure was struck by lightning and the ensuing fire gutted most of the interior The owner at that time rebuilt the cottage replacing almost everything except the outerwalls owned this cottage until she died on May 14 inherited it from her mother and owned it until her death in June inherited the cottage with Courtney eventually becoming the sole owner Her husband was William Peter Van Lopik of Grand Haven Palmer bought Lot 45 in Highland Park in 1912 His purchase included the original cottage Toward the end of the 1910s or sometime in the early 1920s & Kimball Construction firm in Grand Rapids The Owens family probably added to the cottage Owners of the Wickiup in 1999 were Judith M discovered the Grand Haven resort life in 1910 when he came across the lake from Chicago on a steamboat at first staying at bed and breakfast resorts in Highland Park and later buying their own place Mehring's grandparents continued the annual journey to Highland Park until their deaths in the 1930s but their descendants maintained the tradition ever after When Owner Martha Frances Owens died in 1940 she willed the Wickiup to her sister bought the Wickiup in 1948 and came every year until 1982 The cottage may at one time have served as a dormitory for the adjacent Gray Gables since there is evidence that a wide stairway once connected the two cottages Another legend says that it was used as a camp for girls since there are wash basins in three of the bedrooms and a number of initials carved on the porch banister such as the carving on the window frames in the front [Personal correspondence from Judy Mehring.] this summer home was designed and built by Grand Rapids architect David S before having a dispute upset with the Highland Park Association and leaving the area The home was unused and boarded up for several years In 1923 the Woodman family purchased the property Hopkins was designed the Hackley and Hume homes in Muskegon Tom and Mary Ann Donahue added a second story the same year and put in a retaining wall in 1991 Carlie Ringelberg had owned the cottage before Charles Anglin Dirk Buth of Grand Rapids bought this cottage in 1989 from Patricia Dill The 1500 square foot cottage sat on three different ground levels The Ben Lowell family of Grand Haven owned this cottage at least as early as the 1940s and into the 1980s Louis built the residence at the address about 1918 "144" in the cottage's name signifies one gross [a dozen dozen] Cottages on Five Mile Hill once were considered part of Highland Park on the north side of the street and near the top of the hill who called it "Perk's Peak." The Herwyn Inn was originally a boarding house owned and managed by Jane Van Herwyn and her sister The well-known place served wonderful home-cooked meals prepared in the small Marion Blakeslee Smith had fond memories of their romance Ed had formerly owned and managed the concessions at the Pavilion The cottage was passed down and restored by Randy Smith who then sold it to Claudia Kerr and her family This cottage was built by the Goodrich family in 1900 and in the 1950s was in the name of Bernice Goodrich Conklin converted the cottage to a year-round residence A date of 1903 scratched in the woodwork of one of the bedrooms hints at the approximate date when the cottage might have been built Mabel Suter of Canada bought the cottage from the Weber family of St Louis in 1963 was put up by Sarah Benedict Rhines Saunders The cottage remained in its original form until at least 1982 "Loch Hame" was Gaelic for Lake Home Located here was the Thomas Otley summer cottage The Otleys were actively involved in promoting tennis 1414 Lake Avenue [Highland Park Bed and Breakfast] Chicago and St Louis visitors were attracted to the Highland Park Hotel and were soon enjoying the summer breezes and lovely elegance of this hotel built on a high dune overlooking Lake Michigan and nicely appointed rooms drew many guests year after year built as dance hall in 1906 and later converted to suites continued as a bed and breakfast under the ownership of Mell and Ruth Wright who bought the property with the original hotel in 1947 In 1994 Don and Carol Trumbull bought the business from Terry and Judy Postmus John and Sue Smolenski became owners in 1977 The original name of this cottage was "The Sweet Lands." August Boseker built it in 1903 at the same time he constructed three or four others east of here In 1959 it was purchased by a Mr Roddy for $4,250 moved into the cottage and had it completely remodeled and winterized although she retained the original cottage wood 32 Lovers Lane [Beech Holme] [approximate number] built by the Holmes family on Lot 6 of Highland Park plat later was purchased by Fred Darragh in 1919 The residence later was renumbered as 1410 Lake Avenue With two identical sides featuring long side porches and many upstairs bedrooms this cottage was built in 1910 as a duplex The cottage was in Henrietta Warmenhoven's family for at least six generations and contained all the original wicker furniture In 1983 Edward and Sandra Nieuwenhuis of Grand Rapids bought the cottage from Vera Vatthauer's sisters It still had all the original wicker furniture Mary Pearce owned the cottage at this site in the late 1970s Built in 1894 Merry Mac was damaged in a fire before William J The McGrails made some improvements in the cottage but tried to maintain the Highland Park ambience first located at 104 Washington and then 115 Washington In 1989 William and Julie Beaton bought this cottage which had been owned by the family of Herald L The Beatons converted it to a year-round home and occupied by the Hodgekiss family of Joliet when it was bought by Elizabeth Thrall and her family The Thralls also owned the Ship's Lantern [92 Poplar Ridge] This cottage features a large stained glass window in a sitting room adjacent to the living room The gingerbread trim gave it a "Cape Cod" look and feel The wood used in building the cottage was milled on the beach before it was hauled up the hill in order to construct the cottage Illinois was the first owner of the cottage The Hodgekiss family owned a large real estate company and were considered avid golfers It was said that when they brought their cousins and friends to their cottage in the early 1900s a servant in a white coat would greet them on the large porch with prepared drinks awaiting them on a silver tray Isabelle and Marge the two daughters of the Hodgekiss family inherited the cottage and continued to come out to the resort until it the trip became too difficult for them The Applegates acquired ownership in 1946 after selling their share of "Pointe Vue" and "Wrendale" cottages and buying Laquinta The structure of the cottage has not been changed since it was originally completed The cottage displays many fascinating features The unique design allows the front windows to drop down permitting the front of the house to become a screened in porch Another feature of this cottage and many others in Highland Park was the cloth screen that came down over the stairwell to keep the heat downstairs on chilly days when the fireplace was lit The cottage's other unique aspect was the hidden message in its name Five generations of Applegates came to Highland Park for summertime fun so Laquinta held many fond memories for the Applegates Hayes was the most recent owner of this cottage originally named "Ben-Hur" at the time of its building in 1906 Long-time owner Helen Boer became acquainted with the cottage during summer visits to Highland Park Through the years her admiration for the quaint cozy charm of the cottage led to a strong desire to own it Never giving up on her dream she made an offer on the cottage when the opportunity arose in 1934 Although the cottage was remodeled in 1934 the Boers made only minor improvements and Pair-A-Dice was not changed from its original appearance Many of the first owners' furnishings remained in the cottage It was this Helen Boer who wrote Highland Park Yesterday and Today in 1983 Also filling the cottage were family heirlooms of the De Walls such as the chest in the yellow room that belonged to her maternal grandmother More important than the material items that filled the cottage were the cherished memories of the family members and friends who shared in the work and enjoyment of Pair-A-Dice "Ceil Laplage," French for Heaven at the beach occupied by Beryl and Morris Griep from Chicago The Weidemans had been living at 715 Sheldon Road in Grand Haven This small Highland Park cottage boasted originality and comfort with the addition of two decks that expanded into the wooded area behind and the incredible view over Lake Michigan The Weidemans bought the cottage from Morris and Beryl Griep in 1966 The Grieps had bought the cottage from the owners of Dykstra Dry Goods in Grand Rapids in the mid-1950s and took much of the responsibility for taking care of the Tennis Courts In front of this group of cottages there originally was a deck that stretched to a boardwalk connecting with steps which descended to Harbor Drive However the deck deteriorated during World War II but with flamboyant and European decorating completed by Tieke Originally built in 1900 by John and Edna Wagner it was known as the Wagner Hazzard for 25 years who shared it until it was sold to the Grand Rapids De Fouw family who maintained it with much of its natural charm Wigwam was built by the MacNaughton family of Grand Rapids around 1900 In 1979 the original porch was changed in order to take better advantage of the view of Lake Michigan In 1993 the cottage was owned by Sylvia and Walter McNitt The Parks family purchased the lakeside cottage around 1900 Paul Parks and his family remodeled the home to make it suitable for year-round living in 1945   Paul continued to live in the home nearly 100 years later Ship-a-Hoy contains four floors of living space The cottage later was bought by Cliff and Gladys Pfaff Built in the summer of 1899 by George Kornemeyer The cottage changed very little over the years and was still enjoyed by Smedley's descendants five generations later Bill and Bette Waltman were owners of record in 1993 Virginia Johnson Travis owned this cottage Mrs Travis was credited with bringing to Highland Park from China the silver dollar bushes now growing there and throughout the area It was a second home for Grand Haven residents William and Hoppy Herbst for many years   Bert Brouwer and Jane Marshall more recently bought the cottage and maintained its unpretentious and unadorned state Maude Metz and Cora Riggs leased Castle Cranny Crow from Highland Park in 1948 They were teachers from New England who came to Highland Park each summer In 1955 Riggs purchased the cottage from Highland Park and owned it until 1958 when she sold it to the Emersons of Grand Rapids Louis and Genevieve Scott bought it in 1966 and owned it until they sold it to their daughter and son-in-law in 1978 It is believed to have been built in the mid-1890s Known locally as the "Castle," this brick structure was constructed in 1928 to resemble a 15th Century Spanish castle The brick on both the inside and outside of the structure was imported from Italy Various additions were made to the home from time to time the spiral stairway to the widow's watch and the light fixture in the entryway were part of the original structure and were retained through the years In the 1950s and 1960s the Castle served at different times as a restaurant Jerry and Barbara Schuette bought the property in 1999 and spent about $1,000,000 to convert the 6,000 square foot structure to a bed and breakfast the next year They remodeled the third floor to living quarters for themselves making the ballroom into a dining and kitchen area Start Here Resources to help everyone in rowing keep the sport safe and clean and to safeguard participants Safe Sport Resources Competition calendars plus information about entering organising and volunteering at rowing competitions Racing and the popular British Rowing Indoor Championships Indoor Rowing The GB Rowing Team is the high performance arm of British Rowing GB Rowing Team More than 100 people participated in the ROW2025 challenge rowing nearly 4 million metres and raising over £15,0000 for the charity Edwin (front) leads 20 indoor rowers on launch day ROW2025 was a massive community rowing event aiming to raise £2,025 for Kidney Research UK by rowing a cumulative 2025km (2,025,000m) over a week between Saturday 24 January 2025 on multiple machines and on the water Bradford Grammar School BC Captain Edwin van Lopik explained “The challenge is motivated by my brother’s diagnosis of a rare kidney condition last year and I found it particularly difficult to balance supporting my brother with my training and racing aspirations something that really helped us through that time was the incredible support given to us from both of the rowing clubs in Bradford This provided the motivation for ROW2025: to celebrate the strength and unity of Bradford’s small rowing community by doing something incredible for this great cause!” The challenge was launched at Bradford Grammar School (BGS) where 20 rowing machines were kept spinning from 9am until 4.30pm by members of Bradford ARC teachers and parents  all rowing together side-by-side Rowers could also submit the distances they rowed in training later in the week which connected those rowing in Bradford with friends family and club alumni rowing from other locations throughout the UK and Europe The final distance rowed of 3,916,765m was almost twice the original ROW2025 target and included: “To have nearly doubled our distance target is beyond incredible but on top of that this event has reached so many people in such an amazing way and it’s fantastic that we can all row together to reach this distance The fundraising total has also far exceeded everyone’s wildest expectation and is currently standing at nearly £13,000 plus Gift Aid Donate here Special shout outs go to the four people who rowed over 100km during the week: Edwin van Lopik (BGS/BARC) Other notable achievements were 78km from Emma Greenbank (BARC) 37.5km from a BGS Junior Squad member (Y8) and 30km from a BGS pupil who had never rowed before the week began Bradford ARC Captain John Austin-Davies said “Edwin and his brother James are enthusiastic members of Bradford Grammar School Boat Club and are also members of Bradford Amateur Rowing Club Both are determined competitors who put 100% into training We were very concerned when we heard about James’s kidney disorder but it is typical of Ed that he did not sit back but has put together the ROW2025 event to raise funds Bradford ARC is supporting this January Challenge with members from all squads signed up to add kilometres to the total Order yours now British Rowing, 6 Lower Mall, Hammersmith, London, W6 9DJ We\'re sorry, this username is not available, please choose another one 2014 The world’s ‘biggest Christmas tree’ – the 372-metre high transmission mast at Lopik in IJsselstein – will be lit up again this year now the organisers have managed to raise the necessary €65,000 The Lopik tree has been illuminated since 1992 but financial problems mean some years are skipped When the cables stabilising the transmission mast have been hung with lights the tree can be seen up to 30 kilometres away The lights will be switched on December 13 and turned off on January 7 We could not provide the Dutch News service without the generous support of our readers Your donations allow us to report on issues you tell us matter and provide you with a summary of the most important Dutch news each day Many thanks to everyone who has donated to DutchNews.nl in recent days We could not provide this service without you Please help us making DutchNews.nl a better read by taking part in a short survey people all over the world try to make the biggest artificial Christmas tree formed by thousands of lights on the slopes of Mount Ingino Or the illumination of the 372-metre high transmission mast at Lopik in The Netherlands a student of Applied Physics at Delft University of Technology (TU Delft) decided to do the opposite.  She created what is probably the world’s smallest Christmas tree Willems works with a scanning tunneling microscope: a complex device that is capable of scanning individual atoms and even changing their position She uses this microscope to build small structures in order to study their quantum mechanical properties But sometimes you can also use technology for something more fun Willems came up with the idea of making a Christmas tree by removing 51 atoms from a perfect crystal lattice Postbus 5 2600 AA Delft The Netherlands Contact and accessibility Vacancies Reading assistant BrowseAloud Intranet Student portal Donate Disclaimer Privacy & Security Over the past month the Dutch police arrested at least two men suspected of playing major roles in global drug trafficking. A 48-year-old man from Amsterdam was arrested on June 8, and remanded into custody for two months on Monday. And a 48-year-old man was arrested in Rotterdam on Monday, the police announced in statements on Tuesday. The Amsterdam suspect owns several companies in different sectors and is suspected of trafficking drugs and laundering the proceeds thereof for years. The Financial Intelligence Unite received multiple reports of suspicious transactions regarding the suspect and his companies last year. Investigation led the police to believe he had access to several million euros in criminal money. According to the police, the Amsterdam man was directly involved in the shipment of 300 kilograms of hashish, MDMA and amphetamine intercepted in Latvia in August last year. The drugs were found in a truck, the driver is still in custody in Latvia. The man is also believed to own criminal assets and large amounts of drugs is various countries, including 5,400 kilograms of cocaine and over 7 thousand kilos of hashish, the police said. On the day of the man's arrest, the police searched six locations in Amsterdam, Weesp and Lopik and seized two encrypted phones, administration, money, several expensive Rolex watches, a car, multiple properties, and bank accounts. On Tuesday last week, the house and office of a fellow suspect - a 48-year-old man from Amsterdam - were also searched, thought he police did not say whether he was arrested. A 48-year-old man was arrested in Rotterdam on Monday as the police raided nine addresses in the city and one in De Meern. According to the Public Prosecution Service (OM), the man is a leader in the cocaine trafficking world and played a major role in trafficking 1,015 kilograms of cocaine through the port of Antwerp between November 9 and December 21 in 2015, and 3,776 kilos of the drug through the port of Rotterdam between April 14 and June 1 2016. During the raids on Monday, the police seized a Mercedes, several smartphones, valuables, a firearm, at least one PGP encrypted phone, and tens of thousands of euros in cash - including an amount of 15 thousand euros found hidden in a champagne cooler. In the Rotterdam home where the suspect was arrested, the police found many bottles of very expensive drinks, including cognacs and champagnes that sell for several thousand euros per bottle. © 2012-2025, NL Times, All rights reserved. water analysis institute KWR checked the waste water in eight Utrecht towns and villages for cocaine amphetamines and methamphetamine over the period of a week Among the findings: cocaine and cannabis are not as popular as in cities such as Amsterdam and Utrecht but speed is the drug of choice described by the NRC as a closed community where drugs are not mentioned speed is three times more popular than in nearby Utrecht That has been reason enough for local alderman Ad de Regt to draw up a ‘tough plan’ to tackle drugs abuse within the village Amphetamines are also popular in nearby Lopik ‘Speed is farmers’ cocaine,’ Lopik alderman Johan van Everdingen told the NRC ‘It is cheap and easy to get hold of We are going to disrupt the market.’ The EU’s drugs agency also uses waste water analysis to look at drugs consumption throughout Europe Please help us making DutchNews.nl a better read by taking part in a short survey.