Journal
NUCLEIC ACIDS RESEARCH
Volume 36, Issue 15, Pages 5074-5082Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkn489
Keywords
-
Categories
Funding
- Australian Research Council (ARC)
- ARC Linkage (CSIRO)
- Australian Professorial
- Federation Fellowship
- Australian National University
Ask authors/readers for more resources
Escherichia coli DNA polymerase III holoenzyme is composed of 10 different subunits linked by noncovalent interactions. The polymerase activity resides in the-subunit. The-subunit, which contains the proofreading exonuclease site within its N-terminal 185 residues, binds to via a segment of 57 additional C-terminal residues, and also to , whose function is less well defined. The present study shows that greatly enhances the solubility of during cell-free synthesis. In addition, synthesis of in the presence of and resulted in a soluble ternary complex that could readily be purified and analyzed by NMR spectroscopy. Cell-free synthesis of from PCR-amplified DNA coupled with site-directed mutagenesis and selective N-15-labeling provided site-specific assignments of NMR resonances of that were confirmed by lanthanide-induced pseudocontact shifts. The data show that the proofreading domain of is connected to via a flexible linker peptide comprising over 20 residues. This distinguishes the : complex from other proofreading polymerases, which have a more rigid multidomain structure.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available