Dynamics of loading the beta sliding clamp of DNA polymerase III onto DNA.

The Journal of Biological Chemistry
Linda B BloomM F Goodman

Abstract

A "minimal" DNA primer-template system, consisting of an 80-mer template and 30-mer primer, supports processive DNA synthesis by DNA polymerase III core in the presence of the beta sliding clamp, gamma complex clamp loader, and single-stranded binding protein from Escherichia coli. This primer-template system was used to measure the loading of the beta sliding clamp by the gamma complex in an ATP-dependent reaction. Bound protein-DNA complexes were detected by monitoring fluorescence depolarization of DNA. Steady state and time-resolved anisotropies were measured, and stopped-flow pre-steady state fluorescence measurements allowed visualization of the loading reactions in real time. The rate of loading beta onto DNA was 12 s-1, demonstrating that clamp assembly is rapid on the time scale required for lagging strand Okazaki fragment synthesis. The association rate appears to be limited by an intramolecular step occurring prior to the clamp-loading reaction, possibly the opening of the toroidal beta dimer.

References

Dec 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·R H Scheuermann, H Echols
Jan 1, 1995·Annual Review of Biochemistry·Z Kelman, M O'Donnell
Apr 1, 1994·Current Opinion in Genetics & Development·Z Kelman, M O'Donnell

❮ Previous
Next ❯

Citations

Apr 18, 2002·Current Opinion in Structural Biology·David JeruzalmiJohn Kuriyan
Jul 17, 1998·Current Opinion in Chemical Biology·D J SextonS J Benkovic
Aug 26, 1998·Proceedings of the National Academy of Sciences of the United States of America·M TangM F Goodman
Nov 20, 2012·The Journal of Biological Chemistry·Jaclyn N Hayner, Linda B Bloom
Feb 2, 1999·The EMBO Journal·J TurnerM O'Donnell
Nov 26, 2010·Nucleic Acids Research·Michael T MorganAlexander C Drohat
Apr 3, 2013·Cold Spring Harbor Perspectives in Biology·Mark HedglinStephen J Benkovic
Jun 15, 2005·Annual Review of Biochemistry·Aaron Johnson, Mike O'Donnell
Mar 21, 2009·Annual Review of Biochemistry·Samir M Hamdan, Charles C Richardson
Jul 19, 2001·Proceedings of the National Academy of Sciences of the United States of America·M A TrakselisS J Benkovic
Sep 30, 1998·Proceedings of the National Academy of Sciences of the United States of America·J CaiJ Hurwitz
Sep 15, 2004·Journal of Molecular Biology·Maria Magdalena ComanManju M Hingorani
Jun 9, 2006·Critical Reviews in Biochemistry and Molecular Biology·Linda B Bloom
Jun 24, 2004·The Journal of Biological Chemistry·Jeffrey G BertramMyron F Goodman
Feb 13, 2003·The Journal of Biological Chemistry·Aaron Johnson, Mike O'Donnell
Mar 29, 2011·The EMBO Journal·Nathan A TannerAntoine M van Oijen
Apr 13, 2006·Expert Review of Proteomics·Jennifer L BeckNicholas E Dixon
Jun 16, 2006·The Journal of Biological Chemistry·Elvire GuiotEric Deprez
Jan 18, 2014·The Journal of Biological Chemistry·Melissa R MarzahnLinda B Bloom
Dec 10, 1999·The Journal of Biological Chemistry·D M JanzenM K Reddy
Apr 5, 2000·The Journal of Biological Chemistry·X ChenM F Goodman
Sep 2, 2004·The Journal of Biological Chemistry·Brigette TippinMyron F Goodman
Jan 11, 2007·The Journal of Biological Chemistry·Stephen G AndersonLinda B Bloom
Jan 25, 2002·Biophysical Journal·Richard B Dickinson, Daniel L Purich
Jun 20, 1998·Biochemistry·D J SextonS J Benkovic

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.