Nucleic Acids Research Advance Access originally published online on March 13, 2007
Nucleic Acids Research 2007 35(9):2813-2824; doi:10.1093/nar/gkm079
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Nucleic Acids Research, 2007, Vol. 35, No. 9 2813-2824
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Nucleic Acid Enzymes |
The unstructured C-terminus of the 
subunit of Escherichia coli DNA polymerase III holoenzyme is the site of interaction with the 
subunit
1Research School of Chemistry, Australian National University, Canberra ACT 0200, Australia and 2Department of Chemistry, University of Wollongong, NSW 2522, Australia
*To whom correspondence should be addressed. Tel: +61 2 42214346; Fax: +61 2 42214287; E-mail: nick_dixon{at}uow.edu.au
Received October 19, 2006. Revised January 3, 2007. Accepted January 26, 2007.
The 
subunit of Escherichia coli DNA polymerase III holoenzyme interacts with the 
subunit through its C-terminal Domain V, C16. We show that the extreme C-terminal region of C16 constitutes the site of interaction with . The C16 domain, but not a derivative of it with a C-terminal deletion of seven residues (C16
7), forms an isolable complex with . Surface plasmon resonance measurements were used to determine the dissociation constant (KD) of the –C16 complex to be 
260 pM. Competition with immobilized C16 by C16 derivatives for binding to gave values of KD of 7 µM for the –C167 complex. Low-level expression of the genes encoding C16 and C16
7, but not C1611, is lethal to E. coli. Suppression of this lethal phenotype enabled selection of mutations in the 3' end of the C16 gene, that led to defects in binding. The data suggest that the unstructured C-terminus of becomes folded into a helix–loop–helix in its complex with . An N-terminally extended construct, C24, was found to bind DNA in a salt-sensitive manner while no binding was observed for C16, suggesting that the processivity switch of the replisome functionally involves Domain IV of .
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