Structure of the SSB-DNA polymerase III interface and its role in DNA replication Journal Article
| Authors: | Marceau, A. H.; Bahng, S.; Massoni, S. C.; George, N. P.; Sandler, S. J.; Marians, K. J.; Keck, J. L. |
| Article Title: | Structure of the SSB-DNA polymerase III interface and its role in DNA replication |
| 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. © 2011 European Molecular Biology Organization | All Rights Reserved. |
| Keywords: | dna-binding proteins; nonhuman; dna polymerase; dna replication; dna synthesis; protein function; cell cycle; protein protein interaction; protein stability; in vivo study; protein interaction; utilization review; bacterial proteins; amino acid sequence; molecular sequence data; escherichia coli; base sequence; dna structure; dna, single-stranded; biochemistry; escherichia coli proteins; crystallography; replisome; single stranded dna binding protein; temperature dependence; genome maintenance; x-ray crystal structure; dna polymerase iii; holoenzymes |
| Journal Title: | EMBO Journal |
| Volume: | 30 |
| Issue: | 20 |
| ISSN: | 0261-4189 |
| Publisher: | Wiley Blackwell |
| Date Published: | 2011-08-19 |
| Start Page: | 4236 |
| End Page: | 4247 |
| Language: | English |
| DOI: | 10.1038/emboj.2011.305 |
| PROVIDER: | scopus |
| PMCID: | PMC3199393 |
| PUBMED: | 21857649 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 9 December 2011" - "CODEN: EMJOD" - "Source: Scopus" |
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