Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase d-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

Research output: Contribution to journalArticle

25 Citations (Scopus)

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 d 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.

Original languageEnglish (US)
Article numbere22195
JournaleLife
Volume6
DOIs
StatePublished - May 23 2017

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Proliferating Cell Nuclear Antigen
DNA-Directed DNA Polymerase
Annealing
Loss of Heterozygosity
Homologous Recombination
Adenosine Triphosphate

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Liu, J., Ede, C., Wright, W. D., Gore, S. K., Jenkins, S. S., Freudenthal, B. D., ... Heyer, W. D. (2017). Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase d-extending D-loops. eLife, 6, [e22195]. https://doi.org/10.7554/eLife.22195

Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase d-extending D-loops. / Liu, Jie; Ede, Christopher; Wright, William D.; Gore, Steven K.; Jenkins, Shirin S.; Freudenthal, Bret D.; Washington, M. Todd; Veaute, Xavier; Heyer, Wolf Dietrich.

In: eLife, Vol. 6, e22195, 23.05.2017.

Research output: Contribution to journalArticle

Liu, J, Ede, C, Wright, WD, Gore, SK, Jenkins, SS, Freudenthal, BD, Washington, MT, Veaute, X & Heyer, WD 2017, 'Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase d-extending D-loops', eLife, vol. 6, e22195. https://doi.org/10.7554/eLife.22195
Liu, Jie ; Ede, Christopher ; Wright, William D. ; Gore, Steven K. ; Jenkins, Shirin S. ; Freudenthal, Bret D. ; Washington, M. Todd ; Veaute, Xavier ; Heyer, Wolf Dietrich. / Srs2 promotes synthesis-dependent strand annealing by disrupting DNA polymerase d-extending D-loops. In: eLife. 2017 ; Vol. 6.
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