HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 282, Issue 17, 12574-12582, April 27, 2007

This Article Full Text Full Text (PDF) All Versions of this Article: 282/17/12574    most recent M610953200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Pucci, B. Articles by Pisani, F. M. Search for Related Content PubMed PubMed Citation Articles by Pucci, B. Articles by Pisani, F. M. Social Bookmarking                      What's this?

Modular Organization of the Sulfolobus solfataricus Mini-chromosome Maintenance Protein^*

From the Istituto di Biochimica delle Proteine, Consiglio Nazionale Ricerche, Via P. Castellino, 111, 80131 Napoli, Italy

Mini-chromosome maintenance (MCM) proteins form ring-like hexameric complexes that are commonly believed to act as the replicative DNA helicase at the eukaryotic/archaeal DNA replication fork. Because of their simplified composition with respect to the eukaryotic counterparts, the archaeal MCM complexes represent a good model system to use in analyzing the structural/functional relationships of these important replication factors. In this study the domain organization of the MCM-like protein from Sulfolobus solfataricus (Sso MCM) has been dissected by trypsin partial proteolysis. Three truncated derivatives of Sso MCM corresponding to protease-resistant domains were produced as soluble recombinant proteins and purified: the N-terminal domain (N-ter, residues 1–268); a fragment comprising the AAA+ and C-terminal domains (AAA+-C-ter, residues 269–686); and the C-terminal domain (C-ter, residues 504–686). All of the purified recombinant proteins behaved as monomers in solution as determined by analytical gel filtration chromatography, suggesting that the polypeptide chain integrity is required for stable oligomerization of Sso MCM. However, the AAA+-C-ter derivative, which includes the AAA+ motor domain and retains ATPase activity, was able to form dimers in solution when ATP was present, as analyzed by size exclusion chromatography and glycerol gradient sedimentation analyses. Interestingly, the AAA+-C-ter protein could displace oligonucleotides annealed to M13 single-stranded DNA although with a reduced efficiency in comparison with the full-sized Sso MCM. The implications of these findings for understanding the DNA helicase mechanism of the MCM complex are discussed.

Received for publication, November 28, 2006 , and in revised form, March 1, 2007.

This work is dedicated to the memory of Rosa Pucci.

^* This work was supported by a grant from ATIBB-BioTekNet (Centro Regionale di Competenza in Biotecnologie Industriali). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, 80131 Napoli, Italy. Tel.: 39-0816132292; Fax: 39-0816132277; E-mail: fm.pisani{at}ibp.cnr.it.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Samuels, G. Gulati, J.-H. Shin, R. Opara, E. McSweeney, M. Sekedat, S. Long, Z. Kelman, and D. Jeruzalmi A biochemically active MCM-like helicase in Bacillus cereus Nucleic Acids Res., May 27, 2009; (2009) gkp376v1. [Abstract] [Full Text] [PDF]

				A. S. Brewster, G. Wang, X. Yu, W. B. Greenleaf, J. M. Carazo, M. Tjajadi, M. G. Klein, and X. S. Chen Crystal structure of a near-full-length archaeal MCM: Functional insights for an AAA+ hexameric helicase PNAS, December 23, 2008; 105(51): 20191 - 20196. [Abstract] [Full Text] [PDF]

				W. Liu, B. Pucci, M. Rossi, F. M. Pisani, and R. Ladenstein Structural analysis of the Sulfolobus solfataricus MCM protein N-terminal domain Nucleic Acids Res., June 1, 2008; 36(10): 3235 - 3243. [Abstract] [Full Text] [PDF]