Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase δ-extending D-loops

Jie Liu; Christopher Ede; William D Wright; Steven K Gore; Shirin S Jenkins; Bret D Freudenthal; M Todd Washington; Xavier Veaute; Wolf-Dietrich Heyer

doi:10.7554/eLife.22195

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Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase δ-extending D-loops

Journal article

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Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase δ-extending D-loops

Jie Liu, Christopher Ede, William D Wright, Steven K Gore, Shirin S Jenkins, Bret D Freudenthal, M Todd Washington, Xavier Veaute and Wolf-Dietrich Heyer

eLife, Vol.6, e22195

05/23/2017

DOI: 10.7554/eLife.22195

PMCID: PMC5441872

PMID: 28535142

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Abstract

Synthesis-dependent strand annealing (SDSA) is the preferred mode of homologous recombination in somatic cells leading to an obligatory non-crossover outcome, thus avoiding the potential for chromosomal rearrangements and loss of heterozygosity. Genetic analysis identified the Srs2 helicase as a prime candidate to promote SDSA. Here, we demonstrate that Srs2 disrupts D-loops in an ATP-dependent fashion and with a distinct polarity. Specifically, we partly reconstitute the SDSA pathway using Rad51, Rad54, RPA, RFC, DNA Polymerase δ with different forms of PCNA. Consistent with genetic data showing the requirement for SUMO and PCNA binding for the SDSA role of Srs2, Srs2 displays a slight but significant preference to disrupt extending D-loops over unextended D-loops when SUMOylated PCNA is present, compared to unmodified PCNA or monoubiquitinated PCNA. Our data establish a biochemical mechanism for the role of Srs2 in crossover suppression by promoting SDSA through disruption of extended D-loops.

Biochemistry

DNA helicase

genome stability

crossover avoidance

homologous rcombination

Genes and Chromosomes

S. cerevisiae

Details

Title: Subtitle: Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase δ-extending D-loops
Creators: Jie Liu - ,
Christopher Ede - ,
William D Wright - ,
Steven K Gore - ,
Shirin S Jenkins - ,
Bret D Freudenthal - ,
M Todd Washington - ,
Xavier Veaute - ,
Wolf-Dietrich Heyer - ,
Resource Type: Journal article
Publication Details: eLife, Vol.6, e22195
DOI: 10.7554/eLife.22195
PMID: 28535142
PMCID: PMC5441872
NLM abbreviation: Elife
ISSN: 2050-084X
eISSN: 2050-084X
Publisher: eLife Sciences Publications, Ltd
Grant note: ED 162/1-1 / ; CA92276 / ; CA187561 / ; GM081433 / ; GM58015 / ;
Language: English
Date published: 05/23/2017
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025263502771

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