Structural dynamics of E. Coli singlestranded DNA binding protein reveal DNA wrapping and unwrapping pathways

Sukrit Suksombat, Rustem Khafizov, Alexander G. Kozlov, Timothy M. Lohman, Yann R. Chemla

Research output: Contribution to journalArticle

Abstract

Escherichia coli single-stranded (ss)DNA binding (SSB) protein mediates genome maintenance processes by regulating access to ssDNA. This homotetrameric protein wraps ssDNA in multiple distinct binding modes that may be used selectively in different DNA processes, and whose detailed wrapping topologies remain speculative. Here, we used single-molecule force and fluorescence spectroscopy to investigate E. coli SSB binding to ssDNA. Stretching a single ssDNA-SSB complex reveals discrete states that correlate with known binding modes, the likely ssDNA conformations and diffusion dynamics in each, and the kinetic pathways by which the protein wraps ssDNA and is dissociated. The data allow us to construct an energy landscape for the ssDNA-SSB complex, revealing that unwrapping energy costs increase the more ssDNA is unraveled. Our findings provide insights into the mechanism by which proteins gain access to ssDNA bound by SSB, as demonstrated by experiments in which SSB is displa ced by the E. coli recombinase RecA.

Original languageEnglish (US)
Article numbere08193
JournaleLife
Volume4
Issue numberAUGUST2015
DOIs
StatePublished - Aug 25 2015

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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