Rad54 Oligomers Translocate and Cross-bridge Double-stranded DNA to Stimulate Synapsis

Piero R. Bianco, Justin J. Bradfield, Lauren R. Castanza, Andrea N. Donnelly

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Rad54 is a key component of the eukaryotic recombination machinery. Its presence in DNA strand-exchange reactions in vitro results in a significant stimulation of the overall reaction rate. Using untagged Rad54, we show that this stimulation can be attributed to enhancement of the formation of a key reaction intermediate known as DNA networks. Using a novel, single DNA molecule, dual-optical tweezers approach we show how Rad54 stimulates DNA network formation. We discovered that Rad54 oligomers possess a unique ability to cross-bridge or bind double-stranded DNA molecules positioned in close proximity. Further, Rad54 oligomers rapidly translocate double-stranded DNA while simultaneously inducing topological loops in the DNA at the locus of the oligomer. The combination of the cross-bridging and double-stranded DNA translocation activities of Rad54 stimulates the formation of DNA networks, leading to rapid and efficient DNA strand exchange by Rad51.

Original languageEnglish (US)
Pages (from-to)618-640
Number of pages23
JournalJournal of Molecular Biology
Volume374
Issue number3
DOIs
StatePublished - Nov 30 2007
Externally publishedYes

Keywords

  • Chromatin
  • DNA helicase
  • Rad54
  • Recombination
  • Single molecule

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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