Multiple C-terminal tails within a single E coli SSB homotetramer coordinate DNA replication and repair

Edwin Antony, Elizabeth Weiland, Quan Yuan, Carol M. Manhart, Binh Nguyen, Alexander G. Kozlov, Charles S. McHenry, Timothy M. Lohman

Research output: Contribution to journalArticlepeer-review

46 Scopus citations

Abstract

Escherichia coli single-stranded DNA binding protein (SSB) plays essential roles in DNA replication, recombination and repair. SSB functions as a homotetramer with each subunit possessing a DNA binding domain (OB-fold) and an intrinsically disordered C-terminus, of which the last nine amino acids provide the site for interaction with at least a dozen other proteins that function in DNA metabolism. To examine how many C-termini are needed for SSB function, we engineered covalently linked forms of SSB that possess only one or two C-termini within a four-OB-fold "tetramer". Whereas E coli expressing SSB with only two tails can survive, expression of a single-tailed SSB is dominant lethal. E coli expressing only the two-tailed SSB recovers faster from exposure to DNA damaging agents but accumulates more mutations. A single-tailed SSB shows defects in coupled leading and lagging strand DNA replication and does not support replication restart in vitro. These deficiencies in vitro provide a plausible explanation for the lethality observed in vivo. These results indicate that a single SSB tetramer must interact simultaneously with multiple protein partners during some essential roles in genome maintenance.

Original languageEnglish
Pages (from-to)4802-4819
Number of pages18
JournalJournal of Molecular Biology
Volume425
Issue number23
DOIs
StatePublished - Nov 29 2013

Keywords

  • DNA binding
  • DNA repair
  • DNA replication
  • SSB
  • single stranded DNA binding protein

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