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PMID: 10677495 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

A double-hexamer archaeal minichromosome maintenance protein is an ATP-dependent DNA helicase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 4 ·2000-02-15 ·Pages 1530-5

Chong JP, Hayashi MK, Simon MN, Xu RM, Stillman B

Abstract

The minichromosome maintenance (MCM) proteins are essential for DNA replication in eukaryotes. Thus far, all eukaryotes have been shown to contain six highly related MCMs that apparently function together in DNA replication. Sequencing of the entire genome of the thermophilic archaeon Methanobacterium thermoautotrophicum has allowed us to identify only a single MCM-like gene (ORF Mt1770). This gene is most similar to MCM4 in eukaryotic cells. Here we have expressed and purified the M. thermoautotrophicum MCM protein. The purified protein forms a complex that has a molecular mass of approximately 850 kDa, consistent with formation of a double hexamer. The protein has an ATP-independent DNA-binding activity, a DNA-stimulated ATPase activity that discriminates between single- and double-stranded DNA, and a strand-displacement (helicase) activity that can unwind up to 500 base pairs. The 3' to 5' helicase activity requires both ATP hydrolysis and a functional nucleotide-binding site. Moreover, the double hexamer form is the active helicase. It is therefore likely that an MCM complex acts as the replicative DNA helicase in eukaryotes and archaea. The simplified replication machinery in archaea may provide a simplified model for assembly of the machinery required for initiation of eukaryotic DNA replication.

MeSH Terms
Adenosine Triphosphatases/metabolism Adenosine Triphosphate/pharmacology Archaeal Proteins/metabolism,ultrastructure Centrifugation, Density Gradient DNA Helicases/metabolism,ultrastructure DNA Replication DNA-Binding Proteins/metabolism Escherichia coli Magnesium/metabolism Methanobacterium/enzymology Microscopy, Electron Protein Conformation Recombinant Proteins
Chemicals
Archaeal Proteins DNA-Binding Proteins Recombinant Proteins Adenosine Triphosphate Adenosine Triphosphatases DNA Helicases MCM protein, Methanobacterium thermoautotrophicum Magnesium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Chong J P
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
Hayashi M K
Simon M N
Xu R M
Stillman B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.030539597
Published
2000-02-15
Pages
1530-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC26469
Subset
IM
Grants
NCRR NIH HHS · P41 RR001777 · United States
NIGMS NIH HHS · R01 GM045436 · United States
NIGMS NIH HHS · GM45436 · United States
NCRR NIH HHS · P41-RR01777 · United States
External Links
PubMed source DOI: 10.1073/pnas.030539597 PMC Full Text View journal details
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