RecA polymerization on double-stranded DNA by using single-molecule manipulation: The role of ATP hydrolysis

G. V. Shivashankar, M. Feingold, O. Krichevsky, A. Libchaber

Research output: Contribution to journalArticlepeer-review

102 Scopus citations

Abstract

The polymerization of RecA on individual double-stranded DNA molecules is studied. A linear DNA (λ DNA, 48.5 Kb), anchored at one end to a cover glass and at the other end to an optically trapped 3-μm diameter polystyrene bead, serves as a template. The elongation caused by RecA assembly is measured in the presence of ATP and ATP[γS]. By using force extension and hydrodynamic recoil, a value of the persistence length of the RecA-DNA complex is obtained. In the presence of ATP, the polymer length is unstable, first growing to saturation and then decreasing. This suggests a transient dynamics of association and dissociation for RecA on a double-stranded DNA, the process being controlled by ATP hydrolysis. Part of this dynamics is suppressed in the presence of ATP[γS], leading to a stabilized RecA-DNA complex. A one-dimensional nucleation and growth model is presented that may account for the protein assembly.

Original languageEnglish
Pages (from-to)7916-7921
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number14
DOIs
StatePublished - 6 Jul 1999
Externally publishedYes

Keywords

  • DNA-protein interactions
  • Genetic recombination
  • Nucleation and growth
  • Optical tweezers

ASJC Scopus subject areas

  • General

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