PXD036935

PXD036935 is an original dataset announced via ProteomeXchange.

Title	Phase separation properties of RPA combine high-affinity ssDNA binding with dynamic condensate functions at telomeres
Description	RPA has been shown to protect single-stranded DNA (ssDNA) intermediates from instability and breakage. RPA binds ssDNA with sub-nanomolar affinity, yet dynamic turnover is required for downstream ssDNA transactions. How ultrahigh-affinity binding and dynamic turnover are achieved simultaneously is not well understood. Here we reveal that RPA has a strong propensity to assemble into dynamic condensates. In solution, purified RPA phase separates into liquid droplets with fusion and surface wetting behavior. Phase separation is stimulated by sub-stoichiometric amounts of ssDNA, but not RNA or double-stranded DNA, and ssDNA gets selectively enriched in RPA condensates. We find the RPA2 subunit required for condensation and multi-site phosphorylation of the RPA2 N-terminal intrinsically disordered region to regulate RPA self-interaction. Functionally, quantitative proximity proteomics links RPA condensation to telomere clustering and integrity in cancer cells. Collectively, our results suggest that RPA-coated ssDNA is contained in dynamic RPA condensates whose properties are important for genome organization and stability.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:44:12.714.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Vincent Spegg
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-09-22 00:11:03	ID requested
1	2023-03-15 10:24:04	announced
⏵ 2	2023-11-14 08:44:17	announced	2023-11-14: Updated project metadata.

Spegg V, Panagopoulos A, Stout M, Krishnan A, Reginato G, Imhof R, Roschitzki B, Cejka P, Altmeyer M, Phase separation properties of RPA combine high-affinity ssDNA binding with dynamic condensate functions at telomeres. Nat Struct Mol Biol, 30(4):451-462(2023) [pubmed]

submitter keyword: alternative lengthening of telomeres (ALT), replication protein A (RPA),Nuclear organization, biomolecular condensates, genome stability, single-stranded DNA (ssDNA)

Prof. Dr. Matthias Altmeyer
contact affiliation	Department of Molecular Mechanisms of Disease, University of Zurich (UZH), Zurich, Switzerland
contact email	matthias.altmeyer@uzh.ch
lab head
Vincent Spegg
contact affiliation	Department of Molecular Mechanisms of Disease, University of Zurich (UZH), Zurich, Switzerland
contact email	vincent.spegg@dmmd.uzh.ch
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/2023/03/PXD036935

PRIDE project URI

• PRIDE
  1. PXD036935
     1. Label: PRIDE project
     2. Name: Phase separation properties of RPA combine high-affinity ssDNA binding with dynamic condensate functions at telomeres