PXD036486-1

PXD036486 is an original dataset announced via ProteomeXchange.

Title	Mitotic Hela cells expressing GFP-Mitotic Hela cells expressing GFP-Polθ vs Mitotic Hela cells expressing GFP-Polθ treated with Volasertib vs Mitotic Hela cells expressing GFP-Polθ treated with Volasertib
Description	We show that Polθ is recruited to mitotic Double-strand breaks (DSBs) to slow down cell cycle progression and allow DNA repair. Because Polθ is one of the only repair protein to forms repair foci during mitosis, we investigated its regulation during mitosis. We performed immunoprecipitation (IP) of Polθ and assessed phosphorylation by immunoblot analysis (using pan phospho antibodies). We observed a phosphorylation signal corresponding to the size of Polθ when IP was performed from mitotic cell extracts. This phosphorylation signal was abolished when cells where treated with two different PLK1 inhibitors (PLK1i), indicating that PLK1 is responsible for Polθ phosphorylation in mitosis. In order to elucidate the regulation of mitotic Polθ activity, we performed mass spectrometry (MS)-based phosphorylation analysis of Polθ in mitosis with or without the PLK1 inhibitor Volasertib. We found 5 phosphorylated residues. To assess the functional consequences of Polθ phosphorylation by PLK1, we mutated some of the identified residues and found that the phospho dead mutant of Polθ fails be recruited to DSBs in mitosis. This indicates that PLK1-mediated regulation of mitotic Polθ repair is essential for its proper functioning
HostingRepository	PRIDE
AnnounceDate	2023-09-15
AnnouncementXML	Submission_2023-09-15_06:36:34.042.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	VanessaMasson
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-09-02 15:28:38	ID requested
⏵ 1	2023-09-15 06:36:34	announced
2	2023-09-20 08:04:48	announced	2023-09-20: Updated project metadata.
3	2023-09-26 11:06:52	announced	2023-09-26: Updated project metadata.
4	2023-11-14 09:05:25	announced	2023-11-14: Updated project metadata.
5	2024-10-22 06:03:17	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: mitosis, PLK1,Polθ

DamarysLoew
contact affiliation	Head of the Curie Institute Mass Spectrometry Platform
contact email	damarys.loew@curie.Fr
lab head
VanessaMasson
contact affiliation	Institut Curie - Centre de Recherche
contact email	vanessa.masson@curie.fr
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/2023/09/PXD036486

PRIDE project URI

• PRIDE
  1. PXD036486
     1. Label: PRIDE project
     2. Name: Mitotic Hela cells expressing GFP-Mitotic Hela cells expressing GFP-Polθ vs Mitotic Hela cells expressing GFP-Polθ treated with Volasertib vs Mitotic Hela cells expressing GFP-Polθ treated with Volasertib