Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.

<p>(A - B) Western analysis of whole cell extracts from mycATR and mycATRΔC-/- cells. Cells were left untreated (NT) or treated with 0.5 mM of Hydroxyurea (HU) for 20 hours (Chronic) or treated with 5 mM of Hydroxyurea (HU) for 5 hours (Acute), washed and resuspended in fresh media. The sample...

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Príomhchruthaitheoir: Gabriel L. A. da Silva (22676649) (author)
Rannpháirtithe: Jeziel D. Damasceno (9073663) (author), Jennifer A. Black (14411790) (author), Craig Lapsley (6082646) (author), Richard McCulloch (12918) (author), Luiz R. O. Tosi (9073666) (author)
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_version_ 1849927641776259072
author Gabriel L. A. da Silva (22676649)
author2 Jeziel D. Damasceno (9073663)
Jennifer A. Black (14411790)
Craig Lapsley (6082646)
Richard McCulloch (12918)
Luiz R. O. Tosi (9073666)
author2_role author
author
author
author
author
author_facet Gabriel L. A. da Silva (22676649)
Jeziel D. Damasceno (9073663)
Jennifer A. Black (14411790)
Craig Lapsley (6082646)
Richard McCulloch (12918)
Luiz R. O. Tosi (9073666)
author_role author
dc.creator.none.fl_str_mv Gabriel L. A. da Silva (22676649)
Jeziel D. Damasceno (9073663)
Jennifer A. Black (14411790)
Craig Lapsley (6082646)
Richard McCulloch (12918)
Luiz R. O. Tosi (9073666)
dc.date.none.fl_str_mv 2025-11-24T18:33:15Z
dc.identifier.none.fl_str_mv 10.1371/journal.pgen.1011899.g004
dc.relation.none.fl_str_mv https://figshare.com/articles/figure/Deletion_of_the_C-terminus_of_ATR_leads_to_DNA_damage_and_an_increase_of_aberrant_cells_/30697539
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biophysics
Biochemistry
Microbiology
Cell Biology
Genetics
Molecular Biology
Developmental Biology
Science Policy
Infectious Diseases
Biological Sciences not elsewhere classified
Physical Sciences not elsewhere classified
major &# 8217
div >< p
data reveals loss
cells possess mechanisms
remarkably plastic genome
cas9 genome editing
like family acts
protein kinase ataxia
atr depends upon
delayed dna replication
stranded dna accumulation
leishmania major </
dna damage kinase
dna damage
genome stability
leishmania </
dna metabolism
dna injuries
atr acts
replication stress
replication impediments
work shows
whose integrity
terminal knockout
smaller chromosomes
protozoan parasite
phosphatidylinositol 3
nuclear localisation
mycatrδc -/-).
mycatr ),
master regulator
larger chromosomes
key role
increased levels
functional relevance
eukaryotic response
deletion results
constantly challenged
atr plays
atr homolog
atr ),
dc.title.none.fl_str_mv Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
dc.type.none.fl_str_mv Image
Figure
info:eu-repo/semantics/publishedVersion
image
description <p>(A - B) Western analysis of whole cell extracts from mycATR and mycATRΔC-/- cells. Cells were left untreated (NT) or treated with 0.5 mM of Hydroxyurea (HU) for 20 hours (Chronic) or treated with 5 mM of Hydroxyurea (HU) for 5 hours (Acute), washed and resuspended in fresh media. The samples were collected at 0 and 5 hours after chronic treatment, and 0 and 4 hours after acute treatment. Blots were probed with α-γH2A antiserum; α-EIF1α was used as a loading control (predicted protein sizes are indicated, kDa). (C – D) Quantification of signal from the western blot showed in (A and B) using ImageJ software; statistical analysis was conducted using Graph Prism (Error bars ± SD; n = 3 experiments, * p < 0.05; ** p < 0.005; *** p < 0.001; **** p < 0.0001. Unpaired t-test). (E) Representative images acquired by a LSM880 Zeiss confocal microscope from Z stack images of mycATRΔC-/- (cl8) cells after release from chronic treatment (5 hours post treatment). Cells were fixed with 3% formaldehyde, genomic DNA stained with DAPI (cyan) and nucleus (n), kinetoplast (k) and micronucleus (n*) are indicated; α-tubulin was used to stain the cytoskeleton (gray). Bar = 12 um. (F) Percentage of populations with the indicted proportions of Nucleus/Kinetoplast (n/k) in mycATR and mycATRΔC-/- cells untreated (NT) or after release from chronic (5 hours post treatment) or acute (4 hours post treatment) HU treatment. The populations were coloured according to the proportion of n/k as normal (1n1k, 1n2n, 2n2k) or aberrant (0n1k, 1n0k, 2n1k, the presence of a micronucleus, 1M and Others for low aberrant cells as showed 0K2N). Error bars ± SD, n = 2 (>120 cells counted/experiment).</p>
eu_rights_str_mv openAccess
id Manara_71beb741b902baa2e40cf41f0841d33d
identifier_str_mv 10.1371/journal.pgen.1011899.g004
network_acronym_str Manara
network_name_str ManaraRepo
oai_identifier_str oai:figshare.com:article/30697539
publishDate 2025
repository.mail.fl_str_mv
repository.name.fl_str_mv
repository_id_str
rights_invalid_str_mv CC BY 4.0
spelling Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.Gabriel L. A. da Silva (22676649)Jeziel D. Damasceno (9073663)Jennifer A. Black (14411790)Craig Lapsley (6082646)Richard McCulloch (12918)Luiz R. O. Tosi (9073666)BiophysicsBiochemistryMicrobiologyCell BiologyGeneticsMolecular BiologyDevelopmental BiologyScience PolicyInfectious DiseasesBiological Sciences not elsewhere classifiedPhysical Sciences not elsewhere classifiedmajor &# 8217div >< pdata reveals losscells possess mechanismsremarkably plastic genomecas9 genome editinglike family actsprotein kinase ataxiaatr depends upondelayed dna replicationstranded dna accumulationleishmania major </dna damage kinasedna damagegenome stabilityleishmania </dna metabolismdna injuriesatr actsreplication stressreplication impedimentswork showswhose integrityterminal knockoutsmaller chromosomesprotozoan parasitephosphatidylinositol 3nuclear localisationmycatrδc -/-).mycatr ),master regulatorlarger chromosomeskey roleincreased levelsfunctional relevanceeukaryotic responsedeletion resultsconstantly challengedatr playsatr homologatr ),<p>(A - B) Western analysis of whole cell extracts from mycATR and mycATRΔC-/- cells. Cells were left untreated (NT) or treated with 0.5 mM of Hydroxyurea (HU) for 20 hours (Chronic) or treated with 5 mM of Hydroxyurea (HU) for 5 hours (Acute), washed and resuspended in fresh media. The samples were collected at 0 and 5 hours after chronic treatment, and 0 and 4 hours after acute treatment. Blots were probed with α-γH2A antiserum; α-EIF1α was used as a loading control (predicted protein sizes are indicated, kDa). (C – D) Quantification of signal from the western blot showed in (A and B) using ImageJ software; statistical analysis was conducted using Graph Prism (Error bars ± SD; n = 3 experiments, * p < 0.05; ** p < 0.005; *** p < 0.001; **** p < 0.0001. Unpaired t-test). (E) Representative images acquired by a LSM880 Zeiss confocal microscope from Z stack images of mycATRΔC-/- (cl8) cells after release from chronic treatment (5 hours post treatment). Cells were fixed with 3% formaldehyde, genomic DNA stained with DAPI (cyan) and nucleus (n), kinetoplast (k) and micronucleus (n*) are indicated; α-tubulin was used to stain the cytoskeleton (gray). Bar = 12 um. (F) Percentage of populations with the indicted proportions of Nucleus/Kinetoplast (n/k) in mycATR and mycATRΔC-/- cells untreated (NT) or after release from chronic (5 hours post treatment) or acute (4 hours post treatment) HU treatment. The populations were coloured according to the proportion of n/k as normal (1n1k, 1n2n, 2n2k) or aberrant (0n1k, 1n0k, 2n1k, the presence of a micronucleus, 1M and Others for low aberrant cells as showed 0K2N). Error bars ± SD, n = 2 (>120 cells counted/experiment).</p>2025-11-24T18:33:15ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pgen.1011899.g004https://figshare.com/articles/figure/Deletion_of_the_C-terminus_of_ATR_leads_to_DNA_damage_and_an_increase_of_aberrant_cells_/30697539CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/306975392025-11-24T18:33:15Z
spellingShingle Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
Gabriel L. A. da Silva (22676649)
Biophysics
Biochemistry
Microbiology
Cell Biology
Genetics
Molecular Biology
Developmental Biology
Science Policy
Infectious Diseases
Biological Sciences not elsewhere classified
Physical Sciences not elsewhere classified
major &# 8217
div >< p
data reveals loss
cells possess mechanisms
remarkably plastic genome
cas9 genome editing
like family acts
protein kinase ataxia
atr depends upon
delayed dna replication
stranded dna accumulation
leishmania major </
dna damage kinase
dna damage
genome stability
leishmania </
dna metabolism
dna injuries
atr acts
replication stress
replication impediments
work shows
whose integrity
terminal knockout
smaller chromosomes
protozoan parasite
phosphatidylinositol 3
nuclear localisation
mycatrδc -/-).
mycatr ),
master regulator
larger chromosomes
key role
increased levels
functional relevance
eukaryotic response
deletion results
constantly challenged
atr plays
atr homolog
atr ),
status_str publishedVersion
title Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
title_full Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
title_fullStr Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
title_full_unstemmed Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
title_short Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
title_sort Deletion of the C-terminus of ATR leads to DNA damage and an increase of aberrant cells.
topic Biophysics
Biochemistry
Microbiology
Cell Biology
Genetics
Molecular Biology
Developmental Biology
Science Policy
Infectious Diseases
Biological Sciences not elsewhere classified
Physical Sciences not elsewhere classified
major &# 8217
div >< p
data reveals loss
cells possess mechanisms
remarkably plastic genome
cas9 genome editing
like family acts
protein kinase ataxia
atr depends upon
delayed dna replication
stranded dna accumulation
leishmania major </
dna damage kinase
dna damage
genome stability
leishmania </
dna metabolism
dna injuries
atr acts
replication stress
replication impediments
work shows
whose integrity
terminal knockout
smaller chromosomes
protozoan parasite
phosphatidylinositol 3
nuclear localisation
mycatrδc -/-).
mycatr ),
master regulator
larger chromosomes
key role
increased levels
functional relevance
eukaryotic response
deletion results
constantly challenged
atr plays
atr homolog
atr ),