Imaging and energetics of single SSB-ssDNA molecules reveal intramolecular condensation and insight into RecOR function

Jason C. Bell, Bian Liu, Stephen C. Kowalczykowski

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Escherichia coli single-stranded DNA (ssDNA) binding protein (SSB) is the defining bacterial member of ssDNA binding proteins essential for DNA maintenance. SSB binds ssDNA with a variable footprint of ∼30-70 nucleotides, reflecting partial or full wrapping of ssDNA around a tetramer of SSB. We directly imaged single molecules of SSB-coated ssDNA using total internal reflection fluorescence (TIRF) microscopy and observed intramolecular condensation of nucleoprotein complexes exceeding expectations based on simple wrapping transitions. We further examined this unexpected property by single-molecule force spectroscopy using magnetic tweezers. In conditions favoring complete wrapping, SSB engages in long-range reversible intramolecular interactions resulting in condensation of the SSB-ssDNA complex. RecO and RecOR, which interact with SSB, further condensed the complex. Our data support the idea that RecOR-and possibly other SSB-interacting proteins—function(s) in part to alter long-range, macroscopic interactions between or throughout nucleoprotein complexes by microscopically altering wrapping and bridging distant sites.

Original languageEnglish (US)
Article numbere08646
JournaleLife
Volume4
Issue numberSeptember 2015
DOIs
StatePublished - Sep 18 2015

Fingerprint

DNA-Binding Proteins
Condensation
Carrier Proteins
Imaging techniques
Molecules
Nucleoproteins
Single-Stranded DNA
Fluorescence microscopy
Fluorescence Microscopy
Escherichia coli
Nucleotides
Maintenance
Spectroscopy
DNA

ASJC Scopus subject areas

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

Cite this

Imaging and energetics of single SSB-ssDNA molecules reveal intramolecular condensation and insight into RecOR function. / Bell, Jason C.; Liu, Bian; Kowalczykowski, Stephen C.

In: eLife, Vol. 4, No. September 2015, e08646, 18.09.2015.

Research output: Contribution to journalArticle

Bell, Jason C. ; Liu, Bian ; Kowalczykowski, Stephen C. / Imaging and energetics of single SSB-ssDNA molecules reveal intramolecular condensation and insight into RecOR function. In: eLife. 2015 ; Vol. 4, No. September 2015.
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