PXD065573-1

PXD065573 is an original dataset announced via ProteomeXchange.

Title	Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination
Description	Homologous recombination (HR) repairs double-stranded DNA breaks (DSBs). The DSBs are resected to yield single-stranded DNA (ssDNA) that are coated by Replication Protein A (RPA). Rad51 is a recombinase and catalyzes strand invasion and the search for homology. However, it binds to ssDNA with lower affinity than RPA. Thus, mediator proteins such as Rad52 (yeast) or BRCA2 (humans) are required to promote Rad51 binding to RPA-coated ssDNA, but the underlying mechanisms remain poorly understood. Saccharomyces cerevisiae Rad52 interacts with Rad51 through two distinct binding modes. We here uncover that the Rad51-binding site in the disordered C-terminus of Rad52 sorts polydisperse Rad51 into discrete monomers. We developed fluorescent-Rad51 to directly visualize filament formation in single molecule optical tweezer analysis and capture Rad52 catalyzed Rad51 loading onto RPA-coated ssDNA with a distinct preference for junctions. Addition of the Rad51-paralog Rad55-Rad57 increases the number of Rad51 bound by ~60%. Deletion of the C-terminus of Rad52 results in loss of Rad51 sorting and abrogates Rad51 binding to RPA-coated DNA. While BRCA2 and Rad52 are structurally unrelated, we show that many of the functional features are conserved. We describe a concerted Sort, Stack & Extend (SSE) mechanism for mediator-paralog catalyzed Rad51 filament formation in HR.
HostingRepository	PRIDE
AnnounceDate	2025-07-28
AnnouncementXML	Submission_2025-07-27_16:07:13.335.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD065573
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Monika Tokmina-Lukaszewska
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2025-06-27 20:41:57	ID requested
⏵ 1	2025-07-27 16:07:14	announced

10.1038/s41467-025-61811-0;

10.6019/PXD065573;

Deveryshetty J, Mistry A, Pangeni S, Ghoneim M, Tokmina-Lukaszewska M, Gore SK, Liu J, Kaushik V, Karunakaran S, Taddei A, Heyer WD, Ha T, Bothner B, Antony E, Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination. Nat Commun, 16(1):6685(2025) [pubmed]

submitter keyword: cross-linking,LC-MSMS

Edwin Antony
contact affiliation	Department of Biochemistry and Molecular Biology St. Louis University School of Medicine St. Louis, MO 63104, USA
contact email	edwin.antony@health.slu.edu
lab head
Monika Tokmina-Lukaszewska
contact affiliation	Montana State University
contact email	tokminalukas@montana.edu
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/2025/07/PXD065573

PRIDE project URI

• PRIDE
  1. PXD065573
     1. Label: PRIDE project
     2. Name: Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination