Ultrafast redistribution of E. coli SSB along Long single-stranded DNA via intersegment transfer

Kyung Suk Lee, Amanda B. Marciel, Alexander G. Kozlov, Charles M. Schroeder, Timothy M. Lohman, Taekjip Ha

Research output: Contribution to journalArticlepeer-review

Abstract

Single-stranded DNA binding proteins (SSBs) selectively bind single-stranded DNA (ssDNA) and facilitate recruitment of additional proteins and enzymes to their sites of action on DNA. SSB can also locally diffuse on ssDNA, which allows it to quickly reposition itself while remaining bound to ssDNA. In this work, we used a hybrid instrument that combines single-molecule fluorescence and force spectroscopy to directly visualize the movement of Escherichia coli SSB on long polymeric ssDNA. Long ssDNA was synthesized without secondary structure that can hinder quantitative analysis of SSB movement. The apparent diffusion coefficient of E. coli SSB thus determined ranged from 70,000 to 170,000 nt2/s, which is at least 600 times higher than that determined from SSB diffusion on short ssDNA oligomers, and is within the range of values reported for protein diffusion on double-stranded DNA. Our work suggests that SSB can also migrate via a long-range intersegment transfer on long ssDNA. The force dependence of SSB movement on ssDNA further supports this interpretation.

Original languageEnglish (US)
Pages (from-to)2413-2421
Number of pages9
JournalJournal of molecular biology
Volume426
Issue number13
DOIs
StatePublished - Jun 26 2014

Keywords

  • optical tweezer
  • single-stranded DNA
  • single-stranded DNA binding protein

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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