PXD033715

PXD033715 is an original dataset announced via ProteomeXchange.

Title	Microtubule associated proteins MAP7 and MAP7D1 promote 53BP1-mediated DNA double strand break repair in G1 cell cycle phase
Description	In this project we have been looking for proteins binding to phosphorylated sites in DNA repair proteins. Previously, we and others have used peptide-pull down assays as a direct means of attributing specific binding functions to known sites phosphorylated by CK2 in DDR proteins. We have now applied the approach to identify proteins binding to four in vivo phosphorylated sites from DDR proteins RAD50 (Thr690), BRCA1 (Ser1336), MLH1 (Ser477) and XPC (Ser883, 884). These sites are conserved, reside within consensus sites for CK2, share similar sequence features, are mutated in cancer and/or their phosphorylation is known to affect (or is affected) by the DDR. We designed biotinylated 20 amino acid (aa) peptides encompassing the specific phosphorylated amino acids of BRCA1, RAD50, MLH1 and XPC that were either non-phosphorylated or phosphorylated. The biotinylated peptides were coupled to streptavidin beads and used to pull down proteins from HeLa nuclear extracts. Proteomic analysis revealed that all four sites bind proteins in a phospho-dependent manner. Each of the phosphorylated sites were binding various proteins including those implicated in DDR and cell cycle regulation as well as proteins involved in other cellular processes; some of these proteins have already known phospho-binding domains, while others do not. Interestingly, microtubule associated protein MAP7 featured prominently in all of the phosphorylated RAD50, BRCA1, MLH1 and XPC peptide pull-downs and MAP7 paralogue MAP7D1 also featured in the phosphorylated BRCA1 peptide pull-down. In the rest of the project we study the interactions and their function in DNA repair.
HostingRepository	PRIDE
AnnounceDate	2023-03-01
AnnouncementXML	Submission_2023-03-01_10:13:42.961.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD033715
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	VinothiniRajeeve
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue; monohydroxylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-05-05 01:28:39	ID requested
⏵ 1	2023-03-01 10:13:43	announced

10.6019/PXD033715;

submitter keyword: BRCA1,DNA damage repair, proteomics, XPC, G1 cell cycle phase, 53BP1, MAP7, MAP7D1, RAD50, MLH1, LC-MS/MS

Professor Pedro R.Cutillas
contact affiliation	Pedro R. Cutillas Professor of Cell Signalling & Proteomics Centre for Genomics and Computational Biology Barts Cancer Institute - a Cancer Research UK Centre of Excellence Queen Mary, University of London John Vane Science Centre, Charterhouse Square, London EC1M 6BQ
contact email	p.cutillas@qmul.ac.uk
lab head
VinothiniRajeeve
contact affiliation	Centre for Haemato Oncology
contact email	v.rajeeve@qmul.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD033715
     1. Label: PRIDE project
     2. Name: Microtubule associated proteins MAP7 and MAP7D1 promote 53BP1-mediated DNA double strand break repair in G1 cell cycle phase