PMID:21857649
| Citation |
Marceau, AH, Bahng, S, Massoni, SC, George, NP, Sandler, SJ, Marians, KJ and Keck, JL (2011) Structure of the SSB-DNA polymerase III interface and its role in DNA replication. EMBO J. 30:4236-47 |
|---|---|
| Abstract |
Interactions between single-stranded DNA-binding proteins (SSBs) and the DNA replication machinery are found in all organisms, but the roles of these contacts remain poorly defined. In Escherichia coli, SSB's association with the χ subunit of the DNA polymerase III holoenzyme has been proposed to confer stability to the replisome and to aid delivery of primers to the lagging-strand DNA polymerase. Here, the SSB-binding site on χ is identified crystallographically and biochemical and cellular studies delineate the consequences of destabilizing the χ/SSB interface. An essential role for the χ/SSB interaction in lagging-strand primer utilization is not supported. However, sequence changes in χ that block complex formation with SSB lead to salt-dependent uncoupling of leading- and lagging-strand DNA synthesis and to a surprising obstruction of the leading-strand DNA polymerase in vitro, pointing to roles for the χ/SSB complex in replisome establishment and maintenance. Destabilization of the χ/SSB complex in vivo produces cells with temperature-dependent cell cycle defects that appear to arise from replisome instability. |
| Links |
PubMed PMC3199393 Online version:10.1038/emboj.2011.305 |
| Keywords |
Amino Acid Sequence; Bacterial Proteins/chemistry; Bacterial Proteins/metabolism; Base Sequence; DNA Polymerase III/chemistry; DNA Polymerase III/metabolism; DNA Replication; DNA, Single-Stranded/chemistry; DNA, Single-Stranded/metabolism; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/metabolism; Escherichia coli/enzymology; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/metabolism; Holoenzymes/chemistry; Holoenzymes/metabolism; Molecular Sequence Data |
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