PXD035337-2

PXD035337 is an original dataset announced via ProteomeXchange.

Title	Dual role of PCNA-monoubiquitination in facilitating Fanconi Anemia-mediated interstrand crosslink repair
Description	The Fanconi Anemia (FA) repair pathway governs the repair of highly genotoxic DNA interstrand crosslinks (ICLs) with the assistance of translesion synthesis (TLS), that is facilitated by site-specific monoubiquitination of PCNA (PCNA-Ub) at lysine 164 (K164). Mutation at this residue (K164R) renders mammals hypersensitive to ICLs. Besides the FA pathway, alternative pathways have been associated with ICL repair However, the decision-making between the different pathways remains elusive. To study the dependence and relevance of PCNA-Ub in FA repair and pathway preference, we generated a germline Fancg-/- mouse and intercrossed PcnaK164R/+;Fancg-/+ mice. A compound mutation (KR;FGko) was embryonic lethal with the noted exception of very infrequent escapers. RNAseq of primary double mutant mouse embryonic fibroblasts (MEFs) revealed elevated levels of replication stress-induced checkpoints, which were rescued by Trp53 knockdown. Upon crosslink induction, KR and FGko MEFs displayed a similar phenotype regarding sensitivity, cell cycle arrest, fork progression and accumulation of single stranded DNA.Remarkably, PCNA-Ub excludes the mismatch recognition complex MSH2/MSH6 from being recruited. Our results implicate a dual function of PCNA-Ub in ICL repair: 1. Facilitate TLS opposite the unhooked ICL and 2. simultaneously exclude MSH2/MSH6 recruitment to channel the ICL towards canonical FA repair.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_06:53:55.060.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Onno Bleijerveld
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2022-07-14 06:18:09	ID requested
1	2024-08-09 05:22:18	announced
⏵ 2	2024-10-22 06:53:58	announced	2024-10-22: Updated project metadata.

10.1093/pnasnexus/pgae242;

Shah R, Aslam MA, Spanjaard A, de Groot D, Z, ü, rcher LM, Altelaar M, Hoekman L, Pritchard CEJ, Pilzecker B, van den Berk PCM, Jacobs H, Dual role of proliferating cell nuclear antigen monoubiquitination in facilitating Fanconi anemia-mediated interstrand crosslink repair. PNAS Nexus, 3(7):pgae242(2024) [pubmed]

submitter keyword: LC-MSMS, RIME, interstrand crosslink repair, mouse,dna damage tolerance, mismatch repair, fanconi anemia, PCNA ubiquitination

Maarten Altelaar
contact affiliation	The Netherlands Cancer Institute Proteomics Facility, Plesmanlaan 121, 1066CX Amsterdam, The Netherlands; Biomolecular Mass Spectrometry and Proteomics, Utrecht University, The Netherlands
contact email	m.altelaar@nki.nl
lab head
Onno Bleijerveld
contact affiliation	The Netherlands Cancer Institute
contact email	o.bleijerveld@nki.nl
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/08/PXD035337

PRIDE project URI

• PRIDE
  1. PXD035337
     1. Label: PRIDE project
     2. Name: Dual role of PCNA-monoubiquitination in facilitating Fanconi Anemia-mediated interstrand crosslink repair