Proteomic characterization of chromosomal common fragile site (CFS)-associated proteins uncovers ATRX as a regulator of CFS stability.
Chromosome Breakage
Chromosome Fragile Sites
DNA Repair
DNA Replication
/ genetics
Fanconi Anemia
/ genetics
Fanconi Anemia Complementation Group D2 Protein
/ genetics
Genomic Instability
HeLa Cells
Humans
Neoplasms
/ genetics
Proteome
/ genetics
Proteomics
/ methods
RNA Interference
Tandem Mass Spectrometry
X-linked Nuclear Protein
/ genetics
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
05 09 2019
05 09 2019
Historique:
accepted:
30
05
2019
revised:
27
05
2019
received:
18
12
2018
pubmed:
11
6
2019
medline:
18
12
2019
entrez:
11
6
2019
Statut:
ppublish
Résumé
Common fragile sites (CFSs) are conserved genomic regions prone to break under conditions of replication stress (RS). Thus, CFSs are hotspots for rearrangements in cancer and contribute to its chromosomal instability. Here, we have performed a global analysis of proteins that recruit to CFSs upon mild RS to identify novel players in CFS stability. To this end, we performed Chromatin Immunoprecipitation (ChIP) of FANCD2, a protein that localizes specifically to CFSs in G2/M, coupled to mass spectrometry to acquire a CFS interactome. Our strategy was validated by the enrichment of many known regulators of CFS maintenance, including Fanconi Anemia, DNA repair and replication proteins. Among the proteins identified with unknown functions at CFSs was the chromatin remodeler ATRX. Here we demonstrate that ATRX forms foci at a fraction of CFSs upon RS, and that ATRX depletion increases the occurrence of chromosomal breaks, a phenotype further exacerbated under mild RS conditions. Accordingly, ATRX depletion increases the number of 53BP1 bodies and micronuclei, overall indicating that ATRX is required for CFS stability. Overall, our study provides the first proteomic characterization of CFSs as a valuable resource for the identification of novel regulators of CFS stability.
Identifiants
pubmed: 31180492
pii: 5513321
doi: 10.1093/nar/gkz510
pmc: PMC6735892
doi:
Substances chimiques
Fanconi Anemia Complementation Group D2 Protein
0
Proteome
0
ATRX protein, human
EC 3.6.4.12
X-linked Nuclear Protein
EC 3.6.4.12
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
8004-8018Commentaires et corrections
Type : ErratumIn
Informations de copyright
© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.
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