Dynamics and selective remodeling of the DNA-binding domains of RPA

Nilisha Pokhrel; Colleen C Caldwell; Elliot I Corless; Emma A Tillison; Joseph Tibbs; Nina Jocic; S M Ali Tabei; Marc S Wold; Maria Spies; Edwin Antony

doi:10.1038/s41594-018-0181-y

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Dynamics and selective remodeling of the DNA-binding domains of RPA

Journal article

Peer reviewed

Dynamics and selective remodeling of the DNA-binding domains of RPA

Nilisha Pokhrel, Colleen C Caldwell, Elliot I Corless, Emma A Tillison, Joseph Tibbs, Nina Jocic, S M Ali Tabei, Marc S Wold, Maria Spies and Edwin Antony

Nature structural & molecular biology, Vol.26(2), pp.129-136

02/2019

DOI: 10.1038/s41594-018-0181-y

PMCID: PMC6368398

PMID: 30723327

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Abstract

Replication protein A (RPA) coordinates important DNA metabolic events by stabilizing single-stranded DNA (ssDNA) intermediates, activating the DNA-damage response and handing off ssDNA to the appropriate downstream players. Six DNA-binding domains (DBDs) in RPA promote high-affinity binding to ssDNA yet also allow RPA displacement by lower affinity proteins. We generated fluorescent versions of Saccharomyces cerevisiae RPA and visualized the conformational dynamics of individual DBDs in the context of the full-length protein. We show that both DBD-A and DBD-D rapidly bind to and dissociate from ssDNA while RPA remains bound to ssDNA. The recombination mediator protein Rad52 selectively modulates the dynamics of DBD-D. These findings reveal how RPA-interacting proteins with lower ssDNA binding affinities can access the occluded ssDNA and remodel individual DBDs to replace RPA.

Saccharomyces cerevisiae - genetics

DNA, Single-Stranded - metabolism

Humans

Replication Protein A - metabolism

Rad52 DNA Repair and Recombination Protein - genetics

DNA, Single-Stranded - genetics

Saccharomyces cerevisiae - metabolism

Rad52 DNA Repair and Recombination Protein - metabolism

Replication Protein A - chemistry

Protein Binding

Catechin - analogs & derivatives

Catechin - metabolism

Catechin - chemistry

Details

Title: Subtitle: Dynamics and selective remodeling of the DNA-binding domains of RPA
Creators: Nilisha Pokhrel - Department of Biological Sciences, Marquette University, Milwaukee, WI, USA
Colleen C Caldwell - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA, USA
Elliot I Corless - Department of Biological Sciences, Marquette University, Milwaukee, WI, USA
Emma A Tillison - Department of Biological Sciences, Marquette University, Milwaukee, WI, USA
Joseph Tibbs - Department of Physics, University of Northern Iowa, Cedar Falls, IA, USA
Nina Jocic - Department of Physics, University of Northern Iowa, Cedar Falls, IA, USA
S M Ali Tabei - Department of Physics, University of Northern Iowa, Cedar Falls, IA, USA
Marc S Wold - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA, USA
Maria Spies - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA, USA. maria-spies@uiowa.edu
Edwin Antony - Department of Biological Sciences, Marquette University, Milwaukee, WI, USA. edwin.antony@marquette.edu
Resource Type: Journal article
Publication Details: Nature structural & molecular biology, Vol.26(2), pp.129-136
DOI: 10.1038/s41594-018-0181-y
PMID: 30723327
PMCID: PMC6368398
NLM abbreviation: Nat Struct Mol Biol
ISSN: 1545-9993
eISSN: 1545-9985
Publisher: United States
Grant note: R01 GM108617 / NIGMS NIH HHS T32 GM067795 / NIGMS NIH HHS R35 GM131704 / NIGMS NIH HHS R01 GM130746 / NIGMS NIH HHS R15 GM110671 / NIGMS NIH HHS P30 CA086862 / NCI NIH HHS
Language: English
Date published: 02/2019
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025255002771

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