A structural and dynamic model for the assembly of Replication Protein A on single-stranded DNA

Luke A Yates; Ricardo J Aramayo; Nilisha Pokhrel; Colleen C Caldwell; Joshua A Kaplan; Rajika L Perera; Maria Spies; Edwin Antony; Xiaodong Zhang

doi:10.1038/s41467-018-07883-7

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A structural and dynamic model for the assembly of Replication Protein A on single-stranded DNA

Journal article

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A structural and dynamic model for the assembly of Replication Protein A on single-stranded DNA

Luke A Yates, Ricardo J Aramayo, Nilisha Pokhrel, Colleen C Caldwell, Joshua A Kaplan, Rajika L Perera, Maria Spies, Edwin Antony and Xiaodong Zhang

Nature communications, Vol.9(1), pp.5447-14

12/21/2018

DOI: 10.1038/s41467-018-07883-7

PMCID: PMC6303327

PMID: 30575763

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Abstract

Replication Protein A (RPA), the major eukaryotic single stranded DNA-binding protein, binds to exposed ssDNA to protect it from nucleases, participates in a myriad of nucleic acid transactions and coordinates the recruitment of other important players. RPA is a heterotrimer and coats long stretches of single-stranded DNA (ssDNA). The precise molecular architecture of the RPA subunits and its DNA binding domains (DBDs) during assembly is poorly understood. Using cryo electron microscopy we obtained a 3D reconstruction of the RPA trimerisation core bound with ssDNA (∼55 kDa) at ∼4.7 Å resolution and a dimeric RPA assembly on ssDNA. FRET-based solution studies reveal dynamic rearrangements of DBDs during coordinated RPA binding and this activity is regulated by phosphorylation at S178 in RPA70. We present a structural model on how dynamic DBDs promote the cooperative assembly of multiple RPAs on long ssDNA.

Cryoelectron Microscopy

DNA, Single-Stranded - metabolism

Escherichia coli

Fluorescence Resonance Energy Transfer

Models, Structural

Protein Domains

Protein Multimerization

Replication Protein A - genetics

Replication Protein A - metabolism

Saccharomyces cerevisiae

Details

Title: Subtitle: A structural and dynamic model for the assembly of Replication Protein A on single-stranded DNA
Creators: Luke A Yates - Imperial College London
Ricardo J Aramayo - Imperial College London
Nilisha Pokhrel - Marquette University
Colleen C Caldwell - University of Iowa
Joshua A Kaplan - Imperial College London
Rajika L Perera - Imperial College London
Maria Spies - Roy J. and Lucille A. Carver College of Medicine
Edwin Antony - Marquette University
Xiaodong Zhang - Imperial College London
Resource Type: Journal article
Publication Details: Nature communications, Vol.9(1), pp.5447-14
DOI: 10.1038/s41467-018-07883-7
PMID: 30575763
PMCID: PMC6303327
NLM abbreviation: Nat Commun
ISSN: 2041-1723
eISSN: 2041-1723
Grant note: T32 GM067795 / NIGMS NIH HHS Wellcome Trust MR/J001201/1 / Medical Research Council R15 GM110671 / NIGMS NIH HHS P30 CA086862 / NCI NIH HHS
Language: English
Date published: 12/21/2018
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984288722702771

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