Carboxyl-terminal domain III of the delta' subunit of DNA polymerase III holoenzyme binds DnaX and supports cooperative DnaX complex assembly

The Journal of Biological Chemistry
M S Song, C S McHenry

Abstract

The delta' subunit of the DNA polymerase-III holoenzyme is a key component of the DnaX complex; it is required for loading the beta(2) processivity factor onto a primed template. The x-ray crystal structure of delta' indicates a three domain C-shaped structure (Guenther, B., Onrust, R., Sali, A., O'Donnell, M., and Kuriyan, J. (1997) Cell 91, 335-345). In this study, we localized the DnaX-binding domain of delta' to its carboxyl-terminal domain III by quantifying protein-protein interactions using a series of delta' fusion proteins lacking specific domains. The fusion protein corresponding to domain III of delta' bound to DnaX with an affinity approaching that of full-length delta'. In contrast, a construct bearing delta' domains I-II did not bind DnaX at detectable levels. The presence of delta and chi psi strengthened the interaction of DnaX with full-length delta' and delta' domain III. Thus, domain III of delta' not only contains the DnaX-binding site, but also contains the elements required for positive cooperative assembly of the DnaX complex. A domain III-specific anti-delta' monoclonal antibody interfered with DnaX complex formation and abolished the replication activity of DNA polymerase III holoenzyme.

References

Apr 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·Z Tsuchihashi, A Kornberg
May 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·A M Flower, C S McHenry
Jan 1, 1995·Annual Review of Biochemistry·Z Kelman, M O'Donnell
Sep 1, 1995·Molecular and Cellular Biology·G CullmannB Stillman
Jan 11, 1993·Nucleic Acids Research·M O'DonnellJ Hurwitz
Feb 1, 2000·The Journal of Biological Chemistry·B P Glover, C S McHenry
Mar 16, 2000·Proceedings of the National Academy of Sciences of the United States of America·B K Tye
Aug 24, 2001·The Journal of Biological Chemistry·M S SongC S McHenry

❮ Previous
Next ❯

Citations

Dec 26, 2002·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Jun 15, 2005·Annual Review of Biochemistry·Aaron Johnson, Mike O'Donnell
Jun 17, 2011·Annual Review of Biochemistry·Charles S McHenry

❮ 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.