PXD018217-2

PXD018217 is an original dataset announced via ProteomeXchange.

Title	The E3 ubiquitin ligase RFWD3 is required for translesion DNA synthesis
Description	Lesions on DNA can uncouple DNA synthesis from helicase unwinding, generating stretches of unreplicated single stranded DNA (ssDNA) behind the replication fork. These ssDNA gaps need to be filled in to complete DNA duplication and thereby avoid the generation of DNA double-stranded breaks (DSBs), a major source of genomic instability. Gap filling repair involves either translesion DNA synthesis (TLS) or template switching (TS) to bypass the lesion. At the heart of these processes, ubiquitylated PCNA recruits many proteins that dictate pathway choice, but the enzymes regulating PCNA ubiquitylation in vertebrates remain poorly defined. Here we report that the E3 ubiquitin ligase RFWD3 promotes non-specific ubiquitylation of proteins on ssDNA, to enhance protein accumulation and stimulate gap filling repair. The absence of RFWD3 leads to a profound defect in recruitment of key repair and signaling factors to damaged chromatin, including ubiquitin and proteins known to ubiquitylate and interact with ubiquitylated PCNA. As a result, PCNA ubiquitylation is severely inhibited without RFWD3, and TLS across different DNA lesions drastically impaired. We propose that RFWD3 is an essential coordinator of the response to ssDNA gaps, where it triggers wide spread ubiquitylation to drive recruitment of effectors of PCNA ubiquitylation and DNA damage bypass.
HostingRepository	PRIDE
AnnounceDate	2020-12-15
AnnouncementXML	Submission_2021-02-11_22:35:43.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ivo Hendriks
SpeciesList	scientific name: Xenopus laevis (African clawed frog); NCBI TaxID: 8355;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2020-03-26 15:13:19	ID requested
1	2020-12-15 04:18:01	announced
⏵ 2	2021-02-11 22:35:44	announced	2021-02-12: Updated publication reference for PubMed record(s): 33321094.
3	2024-10-22 05:16:42	announced	2024-10-22: Updated project metadata.

Gallina I, Hendriks IA, Hoffmann S, Larsen NB, Johansen J, Colding-Christensen CS, Schubert L, Sell, é, s-Baiget S, F, á, bi, á, n Z, K, ü, hbacher U, Gao AO, R, ä, schle M, Rasmussen S, Nielsen ML, Mailand N, Duxin JP, The ubiquitin ligase RFWD3 is required for translesion DNA synthesis. Mol Cell, 81(3):442-458.e9(2021) [pubmed]

submitter keyword: RFWD3, CHROMASS, Xenopus, ubiquitin, DNA, damage, TLS, TS, PCNA

Michael Lund Nielsen
contact affiliation	Proteomics program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark
contact email	michael.lund.nielsen@cpr.ku.dk
lab head
Ivo Hendriks
contact affiliation	Novo Nordisk Foundation Center for Protein Research, University of Copenhagen
contact email	ivo.a.hendriks@gmail.com
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/2020/12/PXD018217

PRIDE project URI

• PRIDE
  1. PXD018217
     1. Label: PRIDE project
     2. Name: The E3 ubiquitin ligase RFWD3 is required for translesion DNA synthesis