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.
- Authors:
-
- MSKCC
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- FOREIGNNIH
- OSTI Identifier:
- 1033001
- Resource Type:
- Journal Article
- Journal Name:
- EMBO Journal
- Additional Journal Information:
- Journal Volume: 30; Journal Issue: (20) ; 2011; Journal ID: ISSN 0261-4189
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; CELL CYCLE; DEFECTS; DNA; DNA POLYMERASES; DNA REPLICATION; ESCHERICHIA COLI; IN VITRO; IN VIVO; INSTABILITY; MACHINERY; MAINTENANCE; POLYMERASES; PROTEINS; STABILITY; SYNTHESIS
Citation Formats
Marceau, Aimee H, Bahng, Soon, Massoni, Shawn C, George, Nicholas P, Sandler, Steven J, Marians, Kenneth J, Keck, James L, UMASS, Amherst), and UW-MED). Structure of the SSB-DNA polymerase III interface and its role in DNA replication. United States: N. p., 2012.
Web. doi:10.1038/emboj.2011.305.
Marceau, Aimee H, Bahng, Soon, Massoni, Shawn C, George, Nicholas P, Sandler, Steven J, Marians, Kenneth J, Keck, James L, UMASS, Amherst), & UW-MED). Structure of the SSB-DNA polymerase III interface and its role in DNA replication. United States. https://doi.org/10.1038/emboj.2011.305
Marceau, Aimee H, Bahng, Soon, Massoni, Shawn C, George, Nicholas P, Sandler, Steven J, Marians, Kenneth J, Keck, James L, UMASS, Amherst), and UW-MED). 2012.
"Structure of the SSB-DNA polymerase III interface and its role in DNA replication". United States. https://doi.org/10.1038/emboj.2011.305.
@article{osti_1033001,
title = {Structure of the SSB-DNA polymerase III interface and its role in DNA replication},
author = {Marceau, Aimee H and Bahng, Soon and Massoni, Shawn C and George, Nicholas P and Sandler, Steven J and Marians, Kenneth J and Keck, James L and UMASS, Amherst) and UW-MED)},
abstractNote = {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.},
doi = {10.1038/emboj.2011.305},
url = {https://www.osti.gov/biblio/1033001},
journal = {EMBO Journal},
issn = {0261-4189},
number = (20) ; 2011,
volume = 30,
place = {United States},
year = {Tue May 22 00:00:00 EDT 2012},
month = {Tue May 22 00:00:00 EDT 2012}
}