Reconstituting the 4-Strand DNA Strand Exchange

Olga M. Mazina, Alexander V. Mazin

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Proteins of the Rad51 family play a key role in homologous recombination by carrying out DNA strand exchange. Here, we present the methodology and the protocols for the 4-strand exchange between gapped circular DNA and homologous linear duplex DNA promoted by human Rad51 and Escherichia coli RecA orthologs. This reaction includes formation of joint molecules and their extension by branch migration in a polar manner. The presented methodology may be used for reconstitution of the medial-to-late stages of homologous recombination in vitro as well as for investigation of the mechanisms of branch migration by helicase-like proteins, e.g., Rad54, BLM, or RecQ1.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages285-305
Number of pages21
DOIs
StatePublished - 2018
Externally publishedYes

Publication series

NameMethods in Enzymology
Volume600
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Branch migration
  • DNA double-strand break repair
  • DNA strand exchange
  • Holliday junctions
  • Homologous recombination
  • Joint molecules

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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