Proteomics

Dataset Information

0

Phase separation properties of RPA combine high-affinity ssDNA binding with dynamic condensate functions at telomeres


ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Vincent Spegg  

LAB HEAD: Prof. Dr. Matthias Altmeyer

PROVIDER: PXD036935 | Pride | 2023-03-15

REPOSITORIES: Pride

altmetric image

Publications

Phase separation properties of RPA combine high-affinity ssDNA binding with dynamic condensate functions at telomeres.

Spegg Vincent V   Panagopoulos Andreas A   Stout Merula M   Krishnan Aswini A   Reginato Giordano G   Imhof Ralph R   Roschitzki Bernd B   Cejka Petr P   Altmeyer Matthias M  

Nature structural & molecular biology 20230309 4


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  ...[more]

Similar Datasets

2018-07-27 | GSE101536 | GEO
2022-02-22 | PXD028637 | Pride
2018-07-27 | GSE87356 | GEO
2023-10-13 | PXD044158 | Pride
2014-11-20 | E-GEOD-63444 | biostudies-arrayexpress
2011-09-26 | E-GEOD-32416 | biostudies-arrayexpress
2011-09-27 | GSE32416 | GEO
2021-03-01 | GSE149807 | GEO
2014-11-20 | GSE63444 | GEO
2017-01-27 | GSE83648 | GEO