Molecular mechanism of sequence-dependent stability of RecA filament

Sung Hyun Kim, Chirlmin Joo, Taekjip Ha, Doseok Kim

Research output: Contribution to journalArticlepeer-review

Abstract

RecA is a DNA-dependent ATPase and mediates homologous recombination by first forming a filament on a single-stranded (ss) DNA. RecA binds preferentially to TGG repeat sequence, which resembles the recombination hot spot Chi (50-GCTG GTGG-30) and is the most frequent pattern (GTG) of the codon usage in Escherichia coli. Because of the highly dynamic nature of RecA filament formation, which consists of filament nucleation, growth and shrinkage, we need experimental approaches that can resolve each of these processes separately to gain detailed insights into the molecular mechanism of sequence preference. By using a single-molecule fluorescence assay, we examined the effect of sequence on individual stages of nucleation, monomer binding and dissociation. We found that RecA does not recognize the Chi sequence as a nucleation site. In contrast, we observed that it is the reduced monomer dissociation that mainly determines the high filament stability on TGG repeats. This sequence dependence of monomer dissociation is well-correlated with that of ATP hydrolysis, suggesting that DNA sequence dictates filament stability through modulation of ATP hydrolysis.

Original languageEnglish (US)
Pages (from-to)7738-7744
Number of pages7
JournalNucleic acids research
Volume41
Issue number16
DOIs
StatePublished - Sep 2013

ASJC Scopus subject areas

  • Genetics

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