Nucleic Acids Research Advance Access originally published online on January 26, 2007
Nucleic Acids Research 2007 35(3):988-998; doi:10.1093/nar/gkl1117
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Nucleic Acids Research, 2007, Vol. 35, No. 3 988-998
© 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 |
Archaeal MCM has separable processivity, substrate choice and helicase domains
1MRC Cancer Cell Unit, Hutchison MRC Research Centre, Hills Road, Cambridge, CB2 2XZ, UK and 2Center for Advanced Research in Biotechnology, 9600 Gudelsky Dr., Rockville, MD 20850 USA
*To whom correspondence should be addressed. Tel: +44 (0)1223 763311; Fax: +44 (0)1223 763296; E-mail: sb419{at}hutchison-mrc.cam.ac.uk
Received November 10, 2006. Revised November 28, 2006. Accepted December 6, 2006.
The mini-chromosome maintenance (MCM) complex is the principal candidate for the replicative helicase of archaea and eukaryotes. Here, we describe a functional dissection of the roles of the three principal structural modules of the homomultimeric MCM of the hyperthermophilic archaeon Sulfolobus solfataricus. Our results include the first analysis of the central AAA+ domain in isolation. This domain possesses ATPase and helicase activity, defining this as the minimal helicase domain. Reconstitution experiments show that the helicase activity of the AAA+ domain can be stimulated by addition of the isolated N-terminal half in trans. Addition of the N-terminus influences both the processivity of the helicase and the choice of substrate that can be melted by the ATPase domain. The degenerate helix-turn-helix domain at the C-terminus of MCM exerts a negative effect on the helicase activity of the complex. These results provide the first evidence for extensive regulatory inter-domain communication within the MCM complex.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
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