Biochemical Studies on Human Rad51-Mediated Homologous Recombination

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Rad51-mediated pairing between homologous DNA sequences during homologous recombination (HR) plays pivotal roles in DNA double-strand break repair. The multi-step process occurs through cooperation of Rad51 and a number of accessory protein factors. The development of various biochemical analyses with the requisite purified factors provides an opportunity to understand the molecular mechanisms of HR. In this chapter, we describe detailed procedures of in vitro assays using human Rad51, a polypeptide derived from the BRCA2 protein, and the Hop2–Mnd1 complex, to examine (1) homologous DNA pairing, (2) Rad51 targeting to single-stranded DNA, (3) stabilization of the Rad51 nucleoprotein filament, and (4) duplex capture by the Rad51 nucleoprotein filament. These methods are invaluable for delineating the functional interplay of HR factors.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages421-435
Number of pages15
DOIs
StatePublished - 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume745
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • BRCA2
  • DNA pairing
  • homologous
  • homologous recombination
  • Hop2–Mnd1
  • presynaptic filament
  • Rad51

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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