Mechanism of nucleotide discrimination by the translesion synthesis polymerase Rev1

Tyler M Weaver; Timothy H Click; Thu H Khoang; M. Todd Washington; Pratul K Agarwal; Bret D Freudenthal

doi:10.1038/s41467-022-30577-0

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Mechanism of nucleotide discrimination by the translesion synthesis polymerase Rev1

Journal article

Open access

Peer reviewed

Mechanism of nucleotide discrimination by the translesion synthesis polymerase Rev1

Tyler M Weaver, Timothy H Click, Thu H Khoang, M. Todd Washington, Pratul K Agarwal and Bret D Freudenthal

Nature communications, Vol.13(1), pp.2876-2876

05/24/2022

DOI: 10.1038/s41467-022-30577-0

PMCID: PMC9130138

PMID: 35610266

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Abstract

Rev1 is a translesion DNA synthesis (TLS) polymerase involved in the bypass of adducted-guanine bases and abasic sites during DNA replication. During damage bypass, Rev1 utilizes a protein-template mechanism of DNA synthesis, where the templating DNA base is evicted from the Rev1 active site and replaced by an arginine side chain that preferentially binds incoming dCTP. Here, we utilize X-ray crystallography and molecular dynamics simulations to obtain structural insight into the dCTP specificity of Rev1. We show the Rev1 R324 protein-template forms sub-optimal hydrogen bonds with incoming dTTP, dGTP, and dATP that prevents Rev1 from adopting a catalytically competent conformation. Additionally, we show the Rev1 R324 protein-template forms optimal hydrogen bonds with incoming rCTP. However, the incoming rCTP adopts an altered sugar pucker, which prevents the formation of a catalytically competent Rev1 active site. This work provides novel insight into the mechanisms for nucleotide discrimination by the TLS polymerase Rev1.

DNA Repair

DNA Replication

DNA - genetics

DNA - metabolism

DNA Damage

DNA-Directed DNA Polymerase - metabolism

Nucleotides - metabolism

Nucleotidyltransferases - metabolism

Details

Title: Subtitle: Mechanism of nucleotide discrimination by the translesion synthesis polymerase Rev1
Creators: Tyler M Weaver - University of Kansas Medical Center
Timothy H Click - Oklahoma State University
Thu H Khoang - University of Kansas Medical Center
M. Todd Washington - Department of Biochemistry and Molecular Biology, University of Iowa, Iowa City, IA, 52242, USA
Pratul K Agarwal - Oklahoma State University
Bret D Freudenthal - University of Kansas Medical Center
Resource Type: Journal article
Publication Details: Nature communications, Vol.13(1), pp.2876-2876
DOI: 10.1038/s41467-022-30577-0
PMID: 35610266
PMCID: PMC9130138
NLM abbreviation: Nat Commun
ISSN: 2041-1723
eISSN: 2041-1723
Grant note: GM128562 / U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS)
Language: English
Date published: 05/24/2022
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984293083502771

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