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Multiple C-Terminal Tails within a Single E. coli SSB Homotetramer Coordinate DNA Replication and Repair

Author:
Antony, Edwin, Weiland, Elizabeth, Yuan, Quan, Manhart, Carol M., Nguyen, Binh, Kozlov, Alexander G., McHenry, Charles S., Lohman, Timothy M.
Source:
Journal of Molecular Biology 2013 v.425 pp. 4802-4819
ISSN:
0022-2836
Subject:
DNA replication, DNA-binding proteins, Escherichia coli, amino acids, genome, metabolism, mutation, single-stranded DNA
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.
Agid:
1021995