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under the constant loving care of her husband and son
Mary was born in Bakers Summit on January 28
She was the youngest of three children: Billy R
Mary graduated from Roaring Spring High School in 1957
She then attended and graduated from the Altoona Beauty School
Last summer Mary and Bob celebrated 65+ years of marriage
Robert (Bob) Bowser of Osterburg at the Holsinger Church of the Brethren at Bakers Summit
Keith Bowser (Spring Hope) and he married Tracy M
Mary's grandchildren include Shianne N
Mary's great grandchildren include Connor Mayes
age 15 and the apple of her eye twins Jaxton and Jamison Mayes
Mary retired from the Chestnut Ridge School District as a secretary after 39 years
Mary was very active in her church and the Women's Group
Mary was a 42 year member of the Bedford Springs Order of the Eastern Star since November 1983
Mary served multiple positions including past Worthy Matron and served most recently as treasurer
attending sporting events of her grandchildren and great grandchildren
her 2 cats and especially babysitting the twins
Over the years Mary and Bob enjoyed camping from Maine to California to Florida and all parts in between
Her favorite place was Niagara Falls where they spent their 50th anniversary
Mary also enjoyed spending time with her cousins Sally Metzker and Nell Snyder and casino trips with the Ickes family
Burial will be at the convenience of the family
PA 16667 or the Bedford County Humane Society
Online condolences may be expressed at http://www.geiselfuneral.com/
Arrangements entrusted to Geisel-Styer Funeral Homes
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Pennsylvania went to be with the Lord on Wednesday
he married his wife Ruth (Daugherty) Miller
John's United Christ of Church in Bedford
and Everett Seventh-Day Adventist Church in Snake Spring
He worked numerous jobs before retiring from Bedford True Value
from 11 AM to 1PM at Geisel-Styer Funeral Homes
at 1 PM at Geisel-Styer Funeral Homes with Pastor Brad Zembower officiating
Geisel-Styer Funeral Homes & Cremation Services Inc
2024 at her residence. She was born on February 1
1948 in Osterburg a daughter of the late Chester and Dora (Black) Ickes. On August 16
who survives along with a son Corey Bowser and wife Jody
of Schellsburg; and two sisters: Patty Lohr
of Osterburg. She was preceded in death by a brother Larry Ickes
Irma was a teller for Central Bank of Clayburg
and retired as manager after 40 years. She was a member of Trinity Reformed Church in Osterburg. Irma was a avid Pittsburgh Pirates and Steelers fan
Funeral services will be Private. Arrangements by the Timothy A
in Bedford. Our online guestbook is available at www.berkebilefuneralhome.com
OSTERBURG — Hundreds of potential bidders turned out last weekend to not only look for collectibles and bargains
but to say a final farewell to a longtime Bedford County staple
The contents of Slick’s Ivy Stone Restaurant went on the auction block and memorabilia from not only Bedford County but Altoona
Hollidaysburg and surrounding areas was divvied up to the highest bidders
The closing last year of the popular eatery
where the waitresses dressed in Colonial-inspired garb
was a blow to Bedford County and to Osterburg
where the old gas station and diner turned restaurant has stood since 1925
Purchased in 1979 from the Ed Pukala family by the Sam Slick Sr
the restaurant was a stepping stone for many local youngsters getting their first taste of responsibility and a paycheck
In a world where big box stores and chain restaurants seem to rule
the Ivy Stone offered patrons a respite from the modern day as its placement along old Route 220 prevented many folks from using their cellphones in what is considered a “dead zone,” thanks to a curve in the road and tree-covered hillsides
While it was known for its home-cooked meals
new customers would be in awe at the interior
as cocooned within its walls was a treasure trove of artifacts — tin lunch boxes
spinning wheels and a huge assortment of glass juicers
There were also hickory benches hand-built by A.C
The 6-foot long benches were built specifically for the restaurant
who noted that all but two of the benches were purchased at the auction by Latshaw’s descendants
grandfather clocks and crocks joined the lineup alongside a Ronald McDonald doll that drew a lot of interest from the audience — to see who was going to purchase what many said would give them nightmares
with Karns stating everyone he talked to seemed to agree it was “creepy.”
former restaurant employees and customers vying for a piece of history
there were so many items that Karns split auctioneer duties with David Tremmel Jr
and the two would call out and encourage bidders for 30 minutes or so before switching off to rest their voice
A bundt pan brought in $22 as the first item up for bid
but bidders kept their hands down when the restaurant’s piano came up for sale
with no takers for even a single dollar bill
The piano was purchased at auction by his mother
but was functional and used by groups that would reserve dining rooms for Christmas parties and other events
A Blackburn Russell flour sack brought $70
while a grouping of three unnamed flour sacks fetched just $5
showing the crowd was most interested in the local artifacts
and the winning bidders quickly left the venue
apparently having purchased the one item they desired
stayed as the auction went into the afternoon hours
some piling up purchases around their feet
seemingly intent on snagging as many items as they could carry
A metal Planet of the Apes lunch box brought $80
When asked about the sheer number of items
Slick said his mother purchased many of the pieces and they’ve been at the restaurant ever since
Other items were given to the restaurant when local residents came across something they didn’t want
“People just gave us stuff,” Slick and his sister
said as they looked through many of the items up for auction
Easter weekend was an appropriate date for the auction as in year’s past the restaurant would close after Christmas and reopen in time for the spring holiday
The final closing of the restaurant was bittersweet
and despite advertising the business for sale
was “out in the middle of nowhere,” Slick said with a slight shake of his head
Some former residents who now live out of town would come in once or twice a year to visit family
making sure to include a stop at Slick’s in their plans
But the restaurant has stood unused for over a year now
announcing the restaurant would not reopen
“It is with mixed emotions that I write you ‘Goodbye,'” Slick said
I feel that it is not feasible to reopen at this time
To all of you that have made dining with us on Good Friday and Easter Sunday a yearly tradition
“The pandemic started the beginning of the end,” Sam Slick said last weekend
president and CEO of the Bedford County Development Association
said the COVID-19 pandemic “took a heavy toll on several of our restaurants.”
supply chain shortages and difficulty finding labor created huge challenges
“It’s hugely disappointing to see deep-rooted
These businesses have served — and befriended — customers for generations
Slick found himself busy bringing items — mostly Christmas trees and decorations — down from the restaurant’s attic
trying to stay in the background while Hideyo went to the Elizabeth room where the auction took place
she reconnected with customers she got to know as a hostess and waitress
Seeing longtime customers and saying goodbye was hard
“She came back in tears,” he said
adding she knew more of the customers because she worked out front
while he was in the kitchen manning the grill and making his signature dishes — like the Osterburger
a made-to-order burger named after the town and topped with asiago cheese
On a pass through a dining room packed with items not yet sold
Lottie French of Martinsburg reminisced about eating at the restaurant with family
“My favorites were the home fries with onions and gravy,” she said
“My sister’s favorite was the barbecue ribs.”
French said she was looking to purchase a few knick knacks for her walls
something to remember the many times she enjoyed eating at the restaurant
Mark Ruby of Claysburg collects lunch boxes and managed to snag a few for his collection
The former Ivy Stone customer said he also buys and sells collectibles
admitting that he perhaps keeps more than he sells
said he grew up with parents who went to auctions a lot and remembers thinking “I’m never doing this when I grow up.”
he said he has a couple of booths set up at cooperative shops and now has a lot of collectibles
Among his many purchases was an accordion for his girlfriend
but added that she will be able to play the instrument
Some of the items he got will be resold and others he’ll keep for himself
noting he likes to keep the local treasures in the county
the Latshaw benches being purchased by descendants of the craftsman was great
“I’m really glad they stayed in the family,” he said
“That’s really important.”
It was a difficult decision to close the restaurant
but he doesn’t miss cooking for hundreds every day
he can often be found at the Bedford Springs — he’s the one mowing the grass
It’s his second year working at the resort
where he finds mowing relaxing and enjoyable
“It gives me a reason to get up in the morning,” he said with a grin
mentioning golfing on the famed Old Course being one of the perks of the job
he said retiring and selling the restaurant wasn’t taken lightly and he knows how much it meant not only to local residents but to the families who worked there and gathered there for meals
In the Facebook post announcing the closing
he said the Slick “family of employees swelled to well into the hundreds.”
Loyal patrons “made it possible for some of these friends and co-workers to earn enough money to buy their first car
utilities & food bills of so many others in our extended family.”
While he’s not certain of the Ivy Stone’s future now that the unique decor is gone and the restaurant appliances will be up for sale in an online auction
Slick is still hopeful the building will be purchased and put to good use
“It was a nice place for a restaurant,” he said
Antis Township will apply to the state Department of Community and Economic Development for a $250,000 grant to ..
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daughter of the late Clayton and Oleta (Claycomb) Hoover
She is survived by son- Eric husband of Karen (Howard) Waybright
of Bedford; daughter- Marie wife of Lorne Miller
of New Paris; step-son- Mike husband of Trudy (Corle) Waybright
of Alum Bank; step-son- Brian husband of Janice (Allison) Waybright
of Osterburg; step-son- Kevin husband of Robin (Cornell)
grandson- Harmon Waybright; along with 12 step-grandchildren and 12 step-great-grandchildren
Mary is preceded in death by brother- Jim Hoover
and previously worked at Ames Department Store and a bank teller at the Hartley Bank
2024 from 10-11 AM at Geisel-Styer Funeral Homes
Interment will be at Osterburg Community Cemetery
Geisel-Styer Funeral Homes & Cremation Services Inc.
Zelphia is survived by her sons: Allan Yarnall (Karen)
of Bedford; grandchildren: Barry Yarnall (Tammy)
of Rainsburg; Breanna and Brooke Yarnall; several great grandchildren; numerous nieces and nephews and brother-in-law: George Weisel
for 70 years and was a member of the Trinity Reformed Church
She was a member of the Order of the Eastern Start and the "Daughters of Rebekah" Odd fellows Lodge
She will be remembered for the beautiful puzzles she put together and her crocheting skills making lovely clothing and blankets for family and friends
She worked at AW Smith Produce store and was a homemaker
from the hours of 11 AM to 2 PM at the Trinity Reformed Church
A funeral service will be held on Thursday
2024 at 2 PM at the Trinity Reformed Church: 722 Main St
Arrangements entrusted to Geisel-Styer Funeral Home
of Osterburg passed away peacefully on March 3 at James E
PA to Reverend Jesse and Leona (Plummer) Chappell
Dean is survived by two daughters Theresa Horne married to Travis Horne
who he loved like a son; and daughter Jessica Chappell; brother Lloyd Chappell (married to Dana Chappell); sister Linda Weyandt (married to Melvin Weyandt) who both helped take special care of Dean; sister Carol Weyandt (married to Butch Weyandt); grandson Christian Horne who he lovingly referred to as "Pop Pop’s Boy"; and many nephews
he was preceded in death by his siblings Donald Chappell
Dean touched the lives of many with his generous heart
He was a Vietnam veteran who served in the Army from January 1967 through July 1969
During his deployment he served as an equipment engineer
Later in life he was an Over-the-Road truck driver for 35 years hauling many things
Dean enjoyed watching football and westerns
and attending and/or watching church online
A celebration of Dean’s life will be held on Tuesday
March 5th at 6pm at Community Bible Church at 120 West Virginia Road
The service will be co-officiated by Pastor Tim Ellis of Alum Bank God’s Missionary Church and Dean’s nephew Joshua Weyandt. Visitation will be held from 4:30-6pm
Donations can be made in Dean’s memory to Community Bible Church
Military honors will be conducted by the Ft
Online condolences may be expressed at www.akersfuneralhome.com
The family would like to thank the James E
Van Zandt VA Medical Staff on the 5th floor for all their kindness and support
A Thursday night fire in Bedford County left an Osterburg family without a home
The Imler Area Volunteer Fire Company was dispatched about 8:04 p.m
for a dwelling fire on the 1700 block of Heritage Road
The fire at the home of Mark and Denise Lapinski was sparked by a malfunctioning electric space header
Denise Lapinski was home at the time and was alerted to the fire by smoke detectors
though the family’s pet cat was lost in the fire
we would consider the house a total loss,” Dennis said in an email
Serve Pro arrived at the scene later that night and was able to board up the home
The American Red Cross assisted the Lapinskis with temporary housing
About 35 firefighters and emergency medical services personnel assisted on the call
along with the Bedford Area Ambulance Service
Pennsylvania health officials are facing down a potential $500 million loss of federal funding
CLEARFIELD — A Morrisdale man was sentenced in Clearfield County court Monday for assaulting two state troopers ..
The city may apply to the International City and County Management Association for inclusion in a program that ..
A Carrolltown woman is facing nearly 1,500 felony counts after embezzling about $198,500 from her employer over a ..
WASHINGTON — The Federal Reserve will likely keep its key short-term interest rate unchanged on Wednesday
Metrics details
The transcription factor p63 shares a high sequence identity with the tumour suppressor p53 which manifests itself in high structural similarity and preference for DNA sequences
Mutations in the DNA binding domain (DBD) of p53 have been studied in great detail
enabling a general mechanism-based classification
In this study we provide a detailed investigation of all currently known mutations in the p63 DBD
which are associated with developmental syndromes
by measuring their impact on transcriptional activity
zinc binding capacity and thermodynamic stability
Some of the mutations we have further characterized with respect to their ability to convert human dermal fibroblasts into induced keratinocytes
Here we propose a classification of the p63 DBD mutations based on the four different mechanisms of DNA binding impairment which we identified: direct DNA contact
no p63 mutation induces global unfolding and subsequent aggregation of the domain
The dimer interface mutations that affect the DNA binding affinity by disturbing the interaction between the individual DBDs retain partial DNA binding capacity which correlates with a milder patient phenotype
All published case reports of human developmental diseases linked to p63 mutations were extracted and evaluated (Supplementary Table S1)
A An overview of the p63 mutation spectrum was generated by plotting position of all mutations on schematic domain architecture of p63
Each mutation is colour coded according to the predominantly caused developmental disease
B The extracted data was analysed for frequency of mis- and nonsense mutation occurring for each residue in p63 (n = 332 families)
Frameshift mutations and gene deletions were disregarded due to their rare incidence
Mutation frequencies are plotted as bars along the p63 protein sequence aligned to the schematic domain architecture
Each bar is colour coded corresponding to the developmental disease the mutation of the respective residue is predominantly causing
The positions of the six mutational hotspots R204
C Mutation frequency was calculated for the p63 DBD individually (n = 202 families) and blotted with the indicated colour code on the DBD structure (PDB: 3QYN)
The six mutational hotspots are depicted as sticks and highlighted with labels
DNA is shown in light green and the zinc ion as a purple sphere
B TA*: TA*p63 isoform-specific N-terminus; ΔN: ΔNp63 isoform-specific N-terminus; TAD Transactivation domain; DBD DNA binding domain; TD tetramerization domain; SAM sterile-α-motif domain; TID transcriptional inhibitory domain; γ: p63γ isoform-specific C-terminus
nsOFC non‐syndromic orofacial clefting; POI premature ovarian insufficiency; NA not assigned to any of the known mutant p63-related diseases
To obtain a better understanding for the correlation of ELA syndrome characteristics and their underlying molecular impairment of the p63 function we have analysed the so far known p63 mutations in the DBD in detail and investigated their effect on transcription
we have analysed the influence of selected mutations on the conversion of dermal fibroblasts into keratinocyte-like cells by RNA-seq
we focus here only on p63 mutations and compare them to p53
make it likely that the results obtained here for p63 are directly relevant for p73 as well
E DNA binding affinities of purified p63 DBD-TD WT and indicated mutants derived from the fitted binding curves (D)
The bar diagrams shows the equilibrium dissociation constants (KD) with the error bars corresponding to the 95% confidence interval
The bars are coloured in grey whenever an incomplete binding curve was fitted
E Mutations related to ELA syndrome are coloured in blue
mutations related to SHFM4 in green and artificial mutants in brown
R298Q and R280H are still able to bind DNA although with a reduced affinity compared to p63 WT
while the affinities of R280S and R304W are further diminished
The SPR measurements also underscored that the SHFM4 mutation K193E reduces the DNA binding affinity
Overall, the different types of measuring the DNA binding competence have shown that several ELA mutants still bind to DNA, albeit with a reduced affinity, that artificial core domain mutants behave like wild type and that SHFM4 mutations affect DNA binding as well. All results of the in vivo and in vitro DNA binding assays are summarized in Table 1
Mapping of the chemical shift differences onto the structure of the DBD
revealed that the strong structural changes were limited to the zinc finger and its surrounding elements of the DNA binding interface like the loops L2 and L3 that contact the minor groove of the DNA
while the core of the domain remained unperturbed
Mutations related to ELA syndrome are coloured in blue
These data show that loss of Zn binding does not destabilize the DBD core but leads to rearrangement of structural elements important for DNA binding and can thus explain the impaired DNA binding triggered by Zn loss in the H208Y and R204W mutants. All results are summarized in Table 1
Oligomeric states are indicated by ‘m’ (monomer) and ‘d’ (dimer)
while ‘a’ marks high molecular weight species corresponding to aggregates
B Mutations related to ELA syndrome are coloured in blue
Our results so far indicate that most mutations affecting the DNA binding interface directly or inducing a shift to the zinc-free apo state
strongly impair the affinity to DNA and concomitantly the transcriptional activity
the ADULT syndrome mutation R298Q and the SHFM4 mutations K193E and K194E
B–E Mutations related to ELA syndrome are coloured in blue
mutations related to SHFM4 in green and mutations related to AEC/RHS in red
These data indicate that cells expressing p63 R227Q or - to a lesser extent - p63 R298Q maintain most of the function of p63 WT
whereas R304W and R204W mutants are completely impaired
A Schematic representation of the inducible system for conversion of human dermal fibroblasts (HDFs) to keratinocyte-like cells (iKCs)
HDFs with a stable integration of a reverse tetracycline-controlled transactivator (rtTA) are retrovirally transduced with KLF4 only or C-terminally fused to ΔNp63α with a self-cleaving T2A peptide
Conversion is achieved by doxycycline (Doxy)-induced expression of p63 and KLF4 for 72 h
Cells were subsequently analysed by ATAC- and RNA-seq
B Venn diagram of ATAC-seq peaks in HDF expressing either WTp63/KLF4 (light brown) or KLF4 alone (CTR; in light blue) showing the number of open regions (-Log10FDR ≥ 10)
C Percentage of ATAC-seq peak distribution in promoters
or intragenic regulatory elements of the sequences open in both CTR HDF and p63 expressing HDF (cluster 1)
D Upper panel: Heat map representation of the signal density of p63 WT and the indicated DBD mutants sequence tags with a stringency of -Log10FDR ≥ 20
p63 binding sites were distributed within ±5 kb from the WT peak summit
Lower panel: Mean read densities centred (±5 kb) around the peak summits
E Percentage of peak distribution bound by p63 WT or the indicated DBD mutants in promoters
F De novo discovery motifs using STREME performed on 100 bp spanning the summits of the top 1000 ChIP-seq peaks (-Log10FDR ≥ 20) of p63 WT and DBD mutants
G p53 p63 consensus motif scanning (MA0106.30525.1 (TP53)) identified by CentriMo using MEME-ChIP
The DNA binding interface can be divided in two structural elements (Fig. 7A; Supplementary Fig. S1A+B)
organizes the DNA contacting residues R279 and S272
Helix H2 together with the opposing β‐strands S2
S2’ and S10 and loop L1 form the part of the interface
which positions the residues contacting the major groove of the DNA
have not been reported to be mutated in patients
This class of mutations is rather small encompassing only five mutations: G134D, K194E, R227Q and R298Q/G. They impair DNA binding by varying degrees. The mutations do not disturb the zinc finger and are scattered across the DBD (Table 1 and Fig. 7B)
R298 contacts S128 and T130 of the DBD N‐terminus
thereby contributing to its rigid conformation
Loss of the positively charged side chain disrupts the fold of the N‐terminus locally
Introduction of an aspartic acid or valine at the position of G134 has most‐likely a similar effect
R227 lies in the zinc region of the DNA binding interface
but with large distance to the coordinated zinc ion
It does not contribute to the structure of the zinc finger but to the conformation of the loop L2B by interacting with F214 and N215
This effect on the inter-dimer interface is limited to canonical full sites comprised of two conjoined half‐sites without a spacer
because otherwise there is no interaction between the two dimers
The classification described above is based on all currently known mutations
the available number of patient data is relatively small in comparison to mutant p53 data collected from cancer patients
the current absence of certain putative p63 mutations in the literature does not necessarily imply that they cannot occur in p63‐related diseases
the absence of mutations in the hydrophobic core of the p63 DBD correlates with these mutations not affecting p63’s transcriptional activity in our luciferase-based assays
the corresponding p53 residues are cancer mutations that result in global unfolding of the p53 DBD
misregulation of p63 by blocking PTMs could further contribute to the pathogenic mechanism of K193E and K194E and explain their phenotypic association with SHFM4 instead of ELA as DBD mutations
seems to be a mechanism that has more effect on p63 while mutations in the core of the protein and a concomitant destabilization is a mechanism that exists only for p53
the p73 DBD is still significantly more stable than the p53 DBD which most likely prevents mutation-induced global unfolding as seen for p53
The high sequence identity with p63 further suggests that the four mechanisms that impair DNA binding of p63 will apply to p73 as well
All published case reports of human developmental diseases related to p63 mutations were extracted from the PubMed database (as of September 2022) and analysed for mutation distribution and frequency (Supplementary Table 1)
Whenever the number of patients or families carrying a mutation was not clearly stated
the minimal number was assumed for subsequent frequency calculations (recognizable by ‘≥’ in the table) resulting in a total number of 663 patients from 332 families
Each mutation was affiliated with the individual disease it predominantly causes
For transient protein expression in mammalian cells or in-vitro translation of N-terminally Myc-tagged p53 and ΔNp63α
PCR-generated inserts were introduced in pcDNA3.Myc by subcloning using BamHI and XhoI restriction sites yielding pcDNA3.Myc-p53 and pcDNA3.Myc-ΔNp63α
pcDNA3.Myc is a derivative of pcDNA3.1(+) (Thermo Fisher Scientific) engineered with a Myc-tag between HindIII and BamHI sites
All gene inserts lack the intrinsic start codon to avoid expression of untagged proteins via alternative translation initiation skipping the Myc-tag
Any mutations were subsequently introduced by site-directed mutagenesis
adding a new EcoRI site 5’ to the BamHI site and subsequently introducing a Myc-Tag between EcoRI and BamHI sites
The p63 gene insert lacks the intrinsic start codon to avoid expression of untagged proteins via alternative translation initiation skipping the Myc-tag
engineering a self-cleaving T2A peptide (GEGRGSLLTCGDVEENPGPGSG) at the N-terminus via the 5’-oligo
and inserted into the backbone by subcloning using XbaI and NotI restriction sites yielding the final vector
PCR-generated inserts of ΔNp63α wildtype and mutants were introduced by subcloning using XbaI and BamHI restriction sites with the N-terminal Myc-tag added via the 5’-oligo resulting in pRetrox-TRE3G-Myc-ΔNp63α-T2A-KLF4 and the respective mutant p63 derivatives
coli BL21(DE3) Rosetta cells (SGC Frankfurt) were transformed with the respective E
coli expression plasmids for protein production
Cells were grown at 37 °C in 2xYT medium supplemented with 100 µM zinc acetate (Carl Roth) to an optical density (OD) of ~0.8
Protein expression was induced with 1 mM IPTG (Carl Roth) for 16–18 h at 18 °C
cells were grown in LB medium at 37 °C to an OD of ~1
harvested by centrifugation and resuspended in M9 minimal medium supplemented with 100 µM zinc acetate and the appropriate isotopic labelling reagents (1 g/l 15NH4Cl (Cambridge Isotope Laboratories Inc.) and 4 g/l glucose (Carl Roth) for 15N-labelling; 1 g/l 15NH4Cl and 2 g/l 13C-glucose (Cambridge Isotope Laboratories Inc.) for 15N/13C-labelling) to an OD of ~0.3
Cells were then grown at 37 °C to an OD of ~0.8 and protein expression was carried out as described above
After expression cells were harvested by centrifugation and resuspended in IMAC A buffer (25 mM HEPES pH 7.0
20 mM β‐ME,10 μM zinc acetate and 25 mM imidazole) supplemented with DNAse (Sigma-Aldrich)
RNAse (Sigma-Aldrich) and self-prepared protease inhibitor
Cells were lysed by sonification and the lysate was cleared by centrifugation at 4 °C
All proteins were subjected to an initial immobilized metal affinity chromatography (IMAC) purification step
The supernatant was loaded onto HiTrap IMAC Sepharose FF column (Cytiva) pre-equilibrated in IMAC A buffer
bound proteins were washed with 20 column volumes (CV) IMAC A buffer and eluted with 2 CV IMAC B buffer (IMAC A with 300 mM imidazole)
The eluted proteins were supplemented with TEV protease (1:50 w/w; self-prepared) for cleavage of the His-tag and dialysed against IMAC A (DBDs) or IMAC 50 buffer (DBD-TDs; IMAC A buffer supplemented with 50 mM imidazole) over night (ON) at 4 °C
the cleaved tag and any uncleaved protein was removed by a subsequent reverse IMAC step using IMAC A or IMAC 50 buffer
DBD-TDs were further purified by ion-exchange (IEX) chromatography
The salt concentration was reduced below 100 mM by dilution with IEX A buffer (25 mM HEPES pH 7.0
20 mM β‐ME,10 μM zinc acetate) prior loading on HiTrap Heparin HP columns (Cytiva)
Bound protein was eluted with a gradient from 50 mM to 1 M NaCl over 20 CV using IEX B buffer (IEX A with 1 M NaCl)
all proteins were subjected to size exclusion chromatography (SEC) using either a Superdex 75 or Superdex 200 SEC column (Cytiva) equilibrated in assay buffer (25 mM HEPES pH 7.5
0.5 mM TCEP) or NMR buffer (50 mM BIS-TRIS pH 6.8
concentrated by centrifugation (Amicon Ultra Centrifugal Filters
Merck KGaA) and snap frozen in liquid nitrogen for storage at −80 °C until usage
The protocol to prepare the apo form of the p53 and p63 DBD by removing the zinc ion was adapted from Butler et al [50]
Purified DBDs in assay or NMR buffer were treated with 1/33 volume 10% acetic acid and 1/100 volume EDTA (1 M
pH 8) for 5 min on ice and pH was raised again by addition of 1.5 volume HEPES (1 M
The resulting apo DBDs were then transferred in fresh assay or NMR buffer with HiTrap desalting columns (Cytiva)
The specific absorption of the PAR2-Zn2+ complex at 520 nm was measured using Spark multimode microplate reader (Tecan)
30 µM DBDs were incubated with 500 µM zinc binding dye 4-(2-pyridylazo)resorcinol (PAR) (Sigma-Aldrich) and 2.5 mM alkylating agent MMTS (Sigma-Aldrich)
which completely and irreversible releases the zinc ions from cysteine zinc finger proteins like the DBDs
The fraction of DBDs in the zinc‐loaded holo form was determined by calculating the absolute zinc concentration with the help of ZnCl2-PAR standard curve and normalizing it to the protein concentration
30 µM DBDs were incubated with PAR and MMTS
which competes with the zinc finger for the zinc ion
The fraction of dissociated zinc was calculated by normalizing the dissociation sample with PAR only to the respective zinc content sample with PAR and MMTS
Zinc assays were performed in three independent replicates from the same protein purification batch. Statistical significance was assessed by an unpaired t-test or ordinary one-way ANOVA followed by Tukey’s post hoc test using Graphpad Prism 8 (Supplementary Table 3)
Thermal shift assay (TSA) was performed using an iCycler iQ PCR Thermal Cycler (Bio‐Rad) and the SYPRO Orange fluorescent dye (Thermo Fisher Scientific) to determine melting temperatures
purified DBDs were centrifuged at 16,000 xg and 4 °C for 15 min to remove aggregates
Samples were prepared for measurement by mixing 36 μl protein working solution (35 µM in assay buffer) and 4 μl SYPRO Orange working solution (1:200 dilution in assay buffer) in MicroAmp Optical 96‐well plates (Thermo Fisher Scientific)
a thermal gradient from 20 °C to 95 °C with a slope of 1 °C/min was applied and dye fluorescence was recorded every 0.2 °C
The negative first deviation of the fluorescence signal (‐dF/dT) was normalized and smoothed (using Graphpad Prism 8 (GraphPad Software)) before extracting the melting point
Melting temperatures were determined by three independent measurements from the same protein purification batch. Statistical significance was assessed by ordinary one-way ANOVA followed by Tukey’s post hoc test using Graphpad Prism 8 (Supplementary Table 3)
The [1H-15N]-BEST-TROSY HSQC spectrum of the apo DBD was partially assigned based on the spectra of the holo form
After priming the Biacore X‐100 with SPR buffer
the SA chip (Cytiva) was docked and its surface conditioned by three 1 min injections of SPR activation solution (50 mM NaOH and 1 M NaCl) and three 30 s injections of SPR regeneration buffer (25 mM HEPES pH7.5
The dsDNA with the 20 bp p21 RE (DBD-TD) or p63 CS (DBD) was immobilized in flow cell F2 and an equal amount of the dsDNA with the random sequence in flow cell F1
The immobilisation was performed sequentially for the individual flow cells with multiple short injections of the respective dsDNA solution
Unbound DNA was removed by two consecutive 15 s injections of SPR regeneration buffer
An amount of biotinylated dsDNA corresponding to ~100 response units (RU) for the DBD-TD measurements and ~200 RU for the DBD measurements was captured onto the surface of each flow cells
All measurements were performed at 20 °C and in the multi‐cycle format with at least one start‐up cycle followed by nine measurement cycles with increasing concentrations of protein
Each cycle comprised a 3 min injection and association phase followed by a 2 min dissociation phase
Afterwards the surface was regenerated by one or two consecutive 15 s injections of SPR regeneration buffer and washed for 5 min with SPR buffer before starting the next cycle
Measurements were performed consecutively on the same chip
in case of DBD-TD in three replicates of independently prepared dilution series
The non-small cell lung cancer cell line H1299 (ATCC) was used because of its p53 deletion and non-detectable or low endogenous levels of p63 and p73
resulting in low background levels for the functional assays in this study
H1299 cells were cultured in RPMI 1640 medium (Thermo Fisher Scientific) supplemented with 10% Fetal Bovine Serum (FBS
100 U/ml penicillin and 100 µg/ml streptomycin (Thermo Fisher Scientific)
HEK293T cells (a gift from Gian-Paolo Dotto) were used for virus production and cultured in Dulbecco’s in Dulbecco Modified Eagle Medium (Sigma) supplemented with 10% FBS (#ECS5000L
Thermo Fisher Scientific) were cultured in DMEM high glucose with the 10% FBS (#ECS5000L
BJ-HDF were cultured in DMEM-F12 (#ECM0090L
Euroclone) supplemented with either 10% FBS (#ECS5000L
Euroclone) or 10% tetracycline negative FBS (#ECS0182L
All cell lines were cultured at 37 °C and 5% CO2 and were routinely tested for mycoplasma contaminations
H1299 cells were transfected using either the Effectene (Qiagen) or the Lipofectamin 2000 (Thermo Fisher Scientific) transfection reagent according to the manufacturers’ protocols
selected with 2 μg/ml puromycin 48 h after the second infections
and grown for 15 days in the absence of puromycin
Cells from each round of conversion assay were subsequently used for analysis by WB
Each biological replicate of these experiments corresponds to an independent round of infection and passaging
To generate the inducible tetracycline system, the reverse tetracycline-controlled transactivator (rtTA) was introduced via lentivirus Lenti-CMV-rtTA3-Blast (pLenti CMV rtTA3 Blast (w756–1) was a gift from Eric Campeau (Addgene plasmid #26429; http://n2t.net/addgene:26429)) transfected with packaging plasmids into HEK293T cells
Viral suspension was filtered and then supplemented with polybrene (8 mg/ml)
Cells were selected for 1 week with Blasticidin (Thermo Fisher Scientific) at 4 µg/ml
pRetrox-TRE3G-Myc-T2A-KLF4 or pRetrox-TRE3G-Myc-ΔNp63α-T2A-KLF4 wildtype or mutants were used generate retroviruses to infect the BJ-HDF rtTA cell in the presence of polybrene for 2 h at 37 °C and cells were selected using 2 µg/µl puromycin for approximately one week
protein expression in the generated polyclonal stable BJ-HDF cell lines was induced for 72 h using doxycycline (1 µg/ml) which was replenished every 24 h
Proteins were separated by SDS-PAGE using either the XCell SureLock Mini‐Cell SDS‐PAGE system (Thermo Fisher Scientific) or Mini-PROTEAN Tetra Cell SDS‐PAGE system (Bio-Rad) in combination with NuPAGE 4–12%
Bis-Tris Mini Protein gels (Thermo Fisher Scientific) or 4–15% Mini-PROTEAN TGX Stain-Free Precast Protein gels (Bio-Rad)
and subsequently transferred onto PVDF membranes using either the XCell II blot system (Thermo Fisher Scientific) in combination with Immobilon-P transfer membranes (Merck KGaA) or the Trans-Blot Turbo Transfer system (Bio-Rad) in combination with the Trans-Blot Turbo RTA Midi 0.45 µm LF PVDF Transfer Kit (Bio-Rad) according to the manufacturer’s recommendation
Membranes were blocked for 1 h at RT in blocking solution (5% skim milk powder (Sigma-Aldrich) in TBS-t (TBS supplemented with 0.05% (v/v) Tween 20)) and incubated with primary antibody diluted in blocking solution ON at 4 °C
Membranes were washed three times with TBS-t
incubated with the appropriate peroxidase-conjugated secondary antibody (Jackson ImmunoResearch) in blocking solution and wash three times again
Chemiluminescence was detected with Lumi-Imager F1 (Roche) using Amersham ECL Prime WB Detection Reagent (Cytiva)
proteins were separated by SDS-PAGE and blotted onto Immobilon-P transfer membranes (Merck KGaA)
Membranes were blocked with PBS 0.2% Tween 20 with 5% nonfat dry milk and incubated with primary antibodies for 2 h at or ON at 4 °C
After washing three times with PBS 0.2% Tween 20
membranes were incubated for 1 h at RT by using the appropriate horseradish peroxidase-conjugated secondary antibody (Bio-Rad)
Detection was performed by chemiluminescence (Clarity
The following primary antibodies were used in this study: Myc (clone 4A6
Densitometric analysis of western blots was performed using ImageJ (Version 1.51). For the KRT14 protein level of HDF conversion assays, KRT14 signal of each sample was normalized to β-actin. Statistical significance was assessed by ordinary one-way ANOVA followed by Tukey’s post hoc test using Graphpad Prism 8 (Supplementary Table 3)
H1299 cells were harvested 24 h after transfection and lysed for 30 min at RT in 100 µl BN-PAGE lysis buffer (25 mM TRIS-HCl pH 7.5
1 mM DTT and 2 mM MgCl2) supplemented with 1x protease inhibitor (Roche) and 1 µl benzonase (Merck)
Sample were supplemented with 3x BN-PAGE (60% glycerol and 15 mM Coomassie Brilliant Blue G-250) and separated for 1 h at 150 V
using the NativePAGE Bis-Tris Gel System with 3–12% Bis-Tris Mini Protein Gels (Thermo Fisher Scientific) at 4 °C
Subsequent immunoblotting and detection was performed using the XCell II blot system (Thermo Fisher Scientific) as described above with the exception that membranes were destained with methanol and fixed with 8% acetic acid before blocking
lysates were supplemented with 5x SDS-PAGE sample buffer (250 mM TRIS pH 8.0
0.025% (w/v) bromphenol blue sodium salt and 12.5% (v/v) β-ME)
boiled and subjected to SDS-PAGE followed by immunoblotting as described above
cells were fixed in 4% PFA and permeabilized with 0.5% NP40 in PBS
Cells were blocked using 0.5% NP40 in PBS supplied with 5% goat serum and incubated with specific antibody for KRT14 (#905301
Images were acquired using a Leica DMi8 microscope (Leica Microsystems) in combination with the software platform LAS X (Leica Application Suite X
and pcDNA3.1(+) as an empty vector control or pcDNA3.Myc plasmids encoding the indicated Myc-tagged p53 or p63 variants
cells were harvested and resuspended in fresh medium
45 μl cell suspension was transferred into four wells each of a 96-well plate to determine the luciferase signal in technical quadruplicates
To prepare input samples for western blot analysis
resuspended in 100 μl 2x SDS-PAGE sample buffer and boiled
The luciferase reporter assay was performed using the Dual-Glo luciferase assay system (#E2940
Promega) according to the manufacturers’ recommendation
Luminescence signals were measured using a Spark multimode or a GENios Pro microplate reader (Tecan)
The ratio of the Firefly to Renilla luciferase signal was calculated for each technical replicate and the resulting mean of each sample was normalized to the empty vector control and p53 or p63 wildtype to yield the relative activity for each biological replicate
and either pcDNA3.1(+) as an empty vector control
pcDNA3.Myc-p53 alone or pcDNA3.Myc-p53 in combination with increasing amount pcDNA3.Myc plasmids encoding the indicated p63 variants
The total amount of transfected plasmid DNA was kept constant by addition of pcDNA3.1(+) if necessary
the luciferase reporter displacement assay was performed as the conventional variant described above
For all luciferase reporter assays presented in this study at least three independent biological replicates were performed. Statistical significance was assessed by ordinary one-way ANOVA followed by Tukey’s post hoc test using Graphpad Prism 8 (Supplementary Table 3)
100 pmol of biotinylated annealed dsDNA oligonucleotides (Supplementary Table 4) were immobilized on 25 µl Streptavidin Sepharose High Performance affinity resin (Cytiva) in pulldown buffer (50 mM Tris pH 8.0
150 mM NaCl and 0.1% Tween 20) for 1 h at 4 °C
Myc-tagged p63 wildtype and mutants were produced by in-vitro translation (#L1170
Promega) from the respective pcDNA3.Myc plasmids
Reaction were incubated for 90 min at 30 °C and afterwards cleared by centrifugation
5 μl of the reaction were mixed with 95 μl 2x SDS-PAGE sample buffer and boiled
Loaded beads were washed three times with pulldown buffer to remove unbound DNA and incubated with 20 µl in-vitro translated p63 in pulldown buffer with a final volume of 400 µl for 3 h at 4 °C rotating
Bead slurry was transferred into Ultrafree-MC Centrifugal Filter (Merck KGaA) and centrifuged at 250 xg for 1 min
The flow-through was discarded and beads were washed four times with 400 µl ice-cold pulldown buffer by centrifugation
Filters were transferred into a new tubes and bound proteins were eluted by addition of 100 µl boiling 2x SDS-sample buffer followed by centrifugation
Input and pulldown samples were analysed by western blotting and quantified
For the relative pull-down efficiency each pulldown sample was normalized to the input sample and normalized to p63 wildtype. The DNA pull-down assays were performed as three independent biological replicates. Statistical significance was assessed by ordinary one-way ANOVA followed by Tukey’s post hoc test using Graphpad Prism 8 (Supplementary Table 3)
The expression levels of genes were determined with htseq-count 0.9.1 by using cellRanger pre-build genes annotations (Ensembl Assembly 93)
All genes having <1 cpm in less than n_min samples and Perc MM reads >20% simultaneously were filtered out
Differential expression analysis was performed using edgeR5
Genes were filtered using the FDR ≤ 0.01 and gene annotation analysis was performed with Metascape Express analysis
ChIP-seq and ATAC-seq experiments were performed as duplicates in Tet-inducible BJ-HDF cells (see above)
Each replicate corresponds to an independent round of induction of the same set of polyclonal cell lines
4 × 10 ^ 6 cells were cross-linked with 1% formaldehyde in PBS
lysed in 500 μl of SDS lysis Buffer (1% SDS
and sonicated using a Bandelin Sonopuls device in order to obtain DNA fragments of between 200 and 500 bp in size
and supernatant was diluted in ChIP dilution Buffer (0.01% SDS
Samples were then incubated overnight at 4 °C on a rotating wheel with 4 μg of p63-specific antibodies per sample (Abcam
#124762) and 25 μl of Protein A/G Dynabeads (1:1
Beads were washed twice with High Salt Immune Complex Wash Buffer (0.1% SDS
Low Salt Immune Complex Wash Buffer (0.1% SDS
LiCl Immune Complex Wash Buffer (0.25 M LiCl
Elution from the beads was achieved by incubation in elution buffer (1% SDS
100 mM NaHCO3) for 15 min and cross-link was reversed by adding 0.2 M NaCl and incubating overnight at 65 °C
Samples were incubated for 1 h at 45 °C with proteinase K (#EMR023100
EuroClone) and purified using MinElute PCR purification kit (#28004
Fifty-thousand cells were resuspended in lysis buffer (10 mM Tris pH 7.5
0.1% IGEPAL) and centrifuged at 500 x g for 10 min at 4 °C to collect nuclei
Nuclei were placed in 50 μl of Transposase reaction with 2 μl Tagment DNA Enzyme (#20034197
Illumina Tagment DNA Enzyme and Buffer kit
Tagmentation was performed incubating samples at 37 °C for 1 h with 650 rpm shaking
1% SDS and 10 µg/µL Proteinase K) was added to samples and reactions were incubated at 40 °C for 30 min with 650 rpm shaking
DNA purification was performed using AMPure XP for PCR Purification (#A63880
The prepared libraries were then sequenced using the NextSeq 500 (Illumina) according to standard Illumina protocols
Further information on research design is available in the Nature Research Reporting Summary linked to this article
All sequencing data generated during this study have been deposited at GEO (GSE221530)
p63 is essential for regenerative proliferation in limb
p63 is a p53 homologue required for limb and epidermal morphogenesis
Alterations of p63 and p73 in human cancers
Heterozygous germline mutations in the p53 homolog p63 are the cause of EEC syndrome
Isoform-specific roles of mutant p63 in human diseases
Molecular evidence that AEC syndrome and Rapp-Hodgkin syndrome are variable expression of a single genetic disorder
An intermediate phenotype between hay-wells and rapp-hodgkin syndromes in a patient with a novel P63 mutation: confirmation of a variable phenotypic spectrum with a common aetiology
Hay-Wells syndrome is caused by heterozygous missense mutations in the SAM domain of p63
Solution structure of a conserved C-terminal domain of p73 with structural homology to the SAM domain
A C-terminal inhibitory domain controls the activity of p63 by an intramolecular mechanism
Protein aggregation of the p63 transcription factor underlies severe skin fragility in AEC syndrome
Thermodynamic stability of wild-type and mutant p53 core domain
Structural basis for understanding oncogenic p53 mutations and designing rescue drugs
Structure-function-rescue: the diverse nature of common p53 cancer mutants
Quantitative analysis of residual folding and DNA binding in mutant p53 core domain: definition of mutant states for rescue in cancer therapy
Non-oncogenic roles of TAp73: from multiciliogenesis to metabolism
Structural diversity of p63 and p73 isoforms
Cooperative binding of p53 to DNA: regulation by protein-protein interactions through a double salt bridge
High thermostability and lack of cooperative DNA binding distinguish the p63 core domain from the homologous tumor suppressor p53
Conformational stability and activity of p73 require a second helix in the tetramerization domain
and p73: implication for heterotetramer formation
Mechanism of TAp73 inhibition by DeltaNp63 and structural basis of p63/p73 hetero-tetramerization
Pattern of p63 mutations and their phenotypes-update
Intrinsic aggregation propensity of the p63 and p73 TI domains correlates with p53R175H interaction and suggests further significance of aggregation events in the p53 family
Semirational design of active tumor suppressor p53 DNA binding domain with enhanced stability
Adaptive evolution of p53 thermodynamic stability
Highly rapid and efficient conversion of human fibroblasts to keratinocyte-like cells
p63 control of desmosome gene expression and adhesion is compromised in AEC syndrome
A functional enhancer of keratin14 is a direct transcriptional target of deltaNp63
p63 establishes epithelial enhancers at critical craniofacial development genes
Genome-wide analysis of p63 binding sites identifies AP-2 factors as co-regulators of epidermal differentiation
p63 cooperates with CTCF to modulate chromatin architecture in skin keratinocytes
Role of chromatin and transcriptional co-regulators in mediating p63-genome interactions in keratinocytes
Structures of p63 DNA binding domain in complexes with half-site and with spacer-containing full response elements
Pliable DNA conformation of response elements bound to transcription factor p63
The C-terminus of p63 contains multiple regulatory elements with different functions
Altered sumoylation of p63alpha contributes to the split-hand/foot malformation phenotype
Itch/AIP4 associates with and promotes p63 protein degradation
Structure of p73 DNA-binding domain tetramer modulates p73 transactivation
Structure and stability insights into tumour suppressor p53 evolutionary related proteins
The p73 DNA binding domain displays enhanced stability relative to its homologue
scalable generation of high-quality protein multiple sequence alignments using Clustal Omega
A new bioinformatics analysis tools framework at EMBL-EBI
Quality control in oocytes by p63 is based on a spring-loaded activation mechanism on the molecular and cellular level
Advanced mammalian gene-transfer - high titer retroviral vectors with multiple-drug selection markers and a complementary helper-free packaging cell-line
A high-efficiency system for the generation and study of human induced pluripotent stem cells
and aggregation of the zinc-free form of the p53 DNA binding domain
BEST-TROSY experiments for time-efficient sequential resonance assignment of large disordered proteins
Solution structure and binding specificity of the p63 DNA binding domain
Phosphorylation control of p53 DNA-binding cooperativity balances tumorigenesis and aging
Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors
TA*p63 and GTAp63 achieve tighter transcriptional regulation in quality control by converting an inhibitory element into an additional transactivation domain
14-3-3 sigma is a p53-regulated inhibitor of G2/M progression
Mutant p63 affects epidermal cell identity through rewiring the enhancer landscape
seqMINER: an integrated ChIP-seq data interpretation platform
ChIPseeker: an R/Bioconductor package for ChIP peak annotation
BEDTools: a flexible suite of utilities for comparing genomic features
MEME-ChIP: motif analysis of large DNA datasets
Combining evidence using p-values: application to sequence homology searches
JASPAR 2022: the 9th release of the open-access database of transcription factor binding profiles
Download references
We would like to thank TIGEM DNA sequencing facility and Bioinformatics Facility for RNA-seq and initial sequencing and analysis
We like to thank Radboud University RIMLS/Science Faculty sequencing facility for library preparation and sequencing
Funding was provided by the DFG (DO 545/18-1
the Centre for Biomolecular Magnetic Resonance (BMRZ) and the Telethon Foundation (Italy) (GGP20124 to C.M.)
Open Access funding enabled and organized by Projekt DEAL
These authors contributed equally: Christian Osterburg
Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance
CEINGE Biotecnologie Avanzate Franco Salvatore
Radboud Institute of Molecular Life Sciences
Radboud University Nijmegen Medical Centre
Departments of Molecular Developmental Biology
CO and MF conceived the study and designed the experiments
RL and SK performed the experiments and analyzed the data
DA and HZ carried out the ATAC- and ChIP-seq
All authors read and approved the final manuscript
The authors declare no competing interests
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
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DOI: https://doi.org/10.1038/s41419-023-05796-y
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OSTERBURG — This Bedford County community has its very own winter wonderland of Christmas lights and decorations
thanks to the dedication of a family for almost two decades
Chuck Corle has lived in Osterburg his entire life
His father bought more than 30 acres in the ’60s
and different members of the family built houses on separate plots over the years
forming a small community of extended family
live on roughly four acres on Hog Farm Road
The larger pond is home to larger fish like trout
and Chuck often goes fishing in his own private reserve
But when the weather gets colder and the ponds freeze over
the Corles’ property transforms into a miniature version of Lights on the Lake
The Corles set up more than 50 inflatables on the property
as well as hanging lights on the trees and shrubbery
Santa even peeks out of the top floor of the storage shed
What started as a simple setup has evolved into a full-blown attraction each Christmas season
Chuck said the project “started for my grandkids” when they were younger
and although several of them are more grown up now
the lights and blow-ups are still a treasured tradition
Sandy said the first blowup was purchased 18 years ago
There was just the one at first — a snow globe with three snowmen to represent their three grandkids at the time — but more decorations were added each year
After more than 30 years in the Chestnut Ridge School District as a coach and groundskeeper
hunting and fishing have always been some of his favorite hobbies
but the Christmas display has now taken over a lot of that time
but this (commitment) now cuts into that,” Chuck said of the display
It is hard for the couple to enter stores around Christmastime and not leave with another inflatable
They were going to stop with the new additions a few years ago
but they could not help themselves when they saw new blow-ups
we see something new we like,” Chuck said
Chuck said 54 inflatables cover roughly two acres of land
surrounding the ponds and taking up the open space on his lawn
Lights cover his house and the other two buildings on the property
One of the buildings is a storage space for his boats
and the inflatables are stored on the upper level until they are brought back out in November
After almost a year of sitting in the dark and dust
all the inflatables are brought out of the shed and see the light of day around Thanksgiving each year
Chuck and Sandy start the whole process in October with the lights and projectors
but the blow-ups don’t make their appearance until Thanksgiving
One of the extra two buildings on the Corle property houses three breakers for the hookups across the display
The breakers in the house are strictly for the lights on and in the house and the bushes surrounding the house
The Corles’ grandkids still like to participate in the process
often asking to be the ones to flip the breakers each day to see the property “come to life,” Chuck said
The operation has grown so much that drivers can now see the Corles’ property when coming around the bend on Hog Farm Road
The display has become a “local attraction,” Chuck said
but “the parents seem to like it just as much,” he said
Visitors are encouraged to go down the driveway to see all the inflatables up close rather than from the main road
Chuck puts a little bucket out for donations if any visitors feel inclined to help defray the electricity cost — the outside meters can hit up to 300 or 400 kilowatt-hours daily during the week or two leading up to Christmas
Brutally cold weather and winter storms pose a difficult challenge for the Corles
as ice and snow can put a wrinkle in the Christmas display
Chuck said he cleared the inflatables at least three times
He regularly goes around his property and inspects the inflatables and their motors to check for any damage
The inflatables themselves can often be weighed down by snow and ice and won’t stand up properly without some manual adjustments
If wind gusts get as high as predicted today
he will be forced to turn the inflatables off
to let them run throughout the day and night so ice won’t freeze them to the ground
Once he turns the display off for good after New Year’s Day
it only takes a few days to put everything away
even if the stakes that hold down all the inflatables become frozen in the ground
“It takes a month to put everything up
but only days to take down,” Chuck said
Chuck asked Sandy earlier this year if they should take this year off due to the rising electricity costs
but the question was shot down almost immediately
Sandy said the kids would be disappointed if the tradition didn’t continue this year
“I started and now they won’t let me stop,” Chuck said
joking that he will likely only have a year off “when I’m dead.”
The Corles’ property is at 813 Hog Farm Road in Osterburg
The lights and inflatables will be up through New Year’s Day
so there is not much time left to visit the attraction before they are packed up until Thanksgiving 2023
View upcoming auction estimates and receive personalized email alerts for the artists you follow
Pamela Salisbury Gallery is pleased to announce the opening of Lothar Osterburg’s site-wide solo exhibition
The exhibition will continue in the Warren Street galleries through May 12
and in the historic Carriage House through June 9
Lothar Osterburg’s third solo presentation at Pamela Salisbury Gallery
and photogravures from the past twenty-five years
The exhibition continues Osterburg’s reflection on “the artist’s journey”
a theme that began with two earlier shows at PSG, Winterwunderkammer (2021/22) and The Long Way In (2023).
Osterburg remembers his early artist studios with a series of small-scale models nested inside found structures fitted with fisheye lenses
photogravures of the interiors are shown beside each miniature studio
Throughout the remaining five floors of PSG
and models continue his exploration of nautical
Waterline displays an armada of arks at an imaginary waterline suggesting a future sea level
These are paired with a row of small photogravures featuring many of the same boats
photographed over more than twenty-five years
Also on view is work from a series of shipwrecks created in Lake Superior’s Isle Royale National Park
Osterburg staged miniature lighthouses at scenic spots along the shoreline in Acadia National Park—the title taken from a quote on a coastal map on a napkin at a lobster pound.
Osterburg’s fascination with boats and ships dates back to childhood vacations at the North Sea
and he has revisited the subject regularly throughout his career
He built his first models to photograph for his photogravures: three-dimensional boats drawn from bent copper wire
and ships carved from potatoes (which quickly started to dry out)
He has greatly expanded his materials since.
Cecilia Lu’s monstrous-yet-heartwarming assemblages
The tension between optimism and yearning remains taut throughout the artist’s exhibition of photogravures and found-material sculptures
2017 after her third courageous battle with cancer
a daughter of the late Ray and Rose (Hatch) Creighton. On September 11
who survives along with two daughters: Betsie Neff and husband Brad
and Maria Mannion and husband Eric; a son James Ickes and wife Christine
all of Osterburg; six grandchildren: Olivia Lewis
a brother Nick Boyles and wife Mina of Anchorage
She was preceded in death by a sister Dolores "Jean" (Creighton) Christian.
and enjoyed bible studies and fellowship with friends
Grace inspired students through her teaching at the Chestnut Ridge School District for 35 years
and having tea with family and friends.
A Memorial Service will be held on Monday
with Vicar Kathy Popp officiating. Friends and family will be received from 10:00 a.m.
until the hour of the service on Monday at the church
Peter's Lutheran Church in care of Robert Bowser
PA 16667. Arrangements by the Timothy A
of Osterburg went to be with the Lord on Thursday
1940 in King Township a daughter of the late Robert M
and Edythe (Miller) King. On March 20
2005. She is survived by three children: Curtis D
all of Osterburg; six grandchildren: Joshua Weyant and wife Tina
and Morgan and Addison Weyant; and eight great-grandchildren. She was preceded in death by three brothers: Charles
Peggy was a 1958 graduate of Chestnut Ridge High School. She worked as a waitress in the Bedford area for over 30 years
with her most memorable times working at Ed’s Steakhouse
and listening to country and gospel music. Most of all she cherished her children and grandchildren. She will be deeply missed by her family and friends
and will always remain in their hearts. Her family finds comfort in knowing that she has a new body
after a courageous battle with Multiple Sclerosis for over 35 years
and she is now walking with her Lord and Savior
with Pastor David Schoenberger officiating. Burial at Pavia Cemetery. Friends will be received on Tuesday
until the hour of the service. In lieu of flowers contributions can be made to: National Multiple Sclerosis Society; 506 3rd Avenue
PA 16635. Our online guestbook is available at www.berekbilefuneralhome.com
Anyone who knew Linda and Tom can attest that they were “two peas in a pod.” As much as she enjoyed life
she looked forward to seeing her husband again in heaven
Linda was previously married to Gerald “Jerry” J
She tragically lost her first love to cancer in July 1988
Smith (Kelly) of Portage; grandchildren: Dylan-Thomas Stiver
Angelica Smith and Alyssa “Munchkin” Custer; and her siblings (children of Thomas Kinsey Jr.); four great-grandchildren; and a sister
Karen (Beard) Machart in Minnesota; many nieces and nephews; and her beloved Yorkie
She was preceded in death by her siblings: Jane Cruse
Linda attended Hollidaysburg Senior High School and spent her youth attending Foot of Ten Bible Church in Duncansville
She loved being a mom and especially loved art projects
She also spent years as an active member at the Altoona Volunteer Sportsman Club hunting
shooting and enjoying time with her first husband
she enjoyed being a full-time wife and grandmother
She loved taking afternoon motorcycle rides with her husband
Linda was famous with her grandkids for “cinnamon twisties” and spent beach-time with her family in Outer Banks
feeding the deer on her property and Conway Twitty
Linda just enjoyed talking with friends for hours or entire evenings
A celebration of life is being planned for the weekend of June 3
Information on the event will be shared with family and friends via Facebook as the date nears
Online condolences may be expressed at www.jedwardblackburnfh.com
Geisel-Styer Funeral Home & Cremation Services Inc
Graveside Service (Bedford County Memorial Park)
2021 at UPMC Altoona Hospital. She was born on March 21
She was preceded in death by her first husband Guy William Croft and her second husband Robert Gardner. She is survived by four daughters: Beverly A
and Dustin Morgart; eight great-grandchildren; two sisters: Shirley Ickes and husband Don
of Manns Choice; and two brothers: Harry Smith and wife Janet
and Carl “Buck” Smith and wife Carol
of Yellow Creek. She was preceded in death by a son
Croft Sr; and the following siblings: Jeanette Ford
Alice worked at the Everett Sewing Factory for over 20 years and then Walmart before retirement. She raised five children on her own
Her world revolved around her family. Alice loved working in her yard and her flower garden
Funeral services will be private with burial at Fishertown Cemetery. Arrangements by the Timothy A
in Bedford. Funeral services will be live streamed on our Funeral Home Facebook page Friday morning. For flowers and our guestbook please visit www.berkebilefuneralhome.com
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23-year-old Noe Ponti leads a 5-strong Swiss roster headed to Singapore to compete at the 2025 World Championships coming up this summer
Spain repeated as the Men’s Water Polo World Cup champions and secured a spot to compete in Singapore
La decisión tiene efecto inmediato tras una votación en una reunión extraordinaria del European Aquatics Bureau el 24 de abril
we are discussing the madness that occurred at the Ft Lauderdale Pro Swim
Marchand has a few areas of improvement to work on if he hopes to return to his Paris form at the World Championships this summer in Singapore
May 02nd, 2020 Europe, International, News
Six-time European Junior champion Isabel Gose will be joining some of Germany’s top swimmers at SC Magdeburg once practice is able to resume
as the 17-year-old is making the move from SV Nikar Heidelberg “primarily for family reasons.”
“I simply lacked the haven of peace in Heidelberg,” she told DSV, translated from German
“I felt a great need to come back home.”
had moved from Berlin to Osterburg two years ago
which is when Gose departed Potsdamer SC and went to SV Nikar Heidelberg
“I felt very comfortable in Heidelberg too
but it was no longer enough for me to sit on the train for six hours for a short visit to Osterburg,” said the 2019 European medalist
Her new home in Magdeburg is now just a short train ride from Osterburg
two of the best distance swimmers in the world
Kohler broke the women’s short course world record in the 1500 free
Other notable names who have represented SC Magdeburg at recent meets include 2016 European champion Franziska Hentke and Worlds semi-finalist Aliena Schmidtke
Madgeburg is one of a handful of training centers in Germany where a few elite swimmers are being allowed to train at a time
World Champion Philip Heintz is training in Heidelberg
Gose has already found success under the guidance of the head coach in Magdeburg, Bernd Berkhahn
as she followed his training plan for this past SC season
broke the German Record in the 400 free in November
and followed that up by winning silver at the European Championships in the event in December
Gose also won a bronze medal at the 2018 European Long Course Championships in the women’s 4×200 free relay
At the European Junior Championships she owns a staggering 14 medals from her two appearances
including a 2018 gold in the 200 free and five titles in 2019 (100
women’s and mixed 4×100 free relay)
SC Magdeburg is a multi-sport club in Germany that offers handball
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