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

James P J Chong, Mariko K. Hayashi, Martha N. Simon, Rui Ming Xu, Bruce Stillman

Research output: Contribution to journalArticle

248 Citations (Scopus)

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 ≃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.

Original languageEnglish
Pages (from-to)1530-1535
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume97
Issue number4
DOIs
Publication statusPublished - 2000 Feb 15
Externally publishedYes

Fingerprint

Archaeal Proteins
Minichromosome Maintenance Proteins
DNA Helicases
Archaea
Eukaryota
DNA Replication
DNA
Adenosine Triphosphate
Maintenance
Methanobacterium
Single-Stranded DNA
Eukaryotic Cells
Base Pairing
Open Reading Frames
Genes
Adenosine Triphosphatases
Proteins
Hydrolysis
Nucleotides
Binding Sites

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

A double-hexamer archaeal minichromosome maintenance protein is an ATP- dependent DNA helicase. / Chong, James P J; Hayashi, Mariko K.; Simon, Martha N.; Xu, Rui Ming; Stillman, Bruce.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, No. 4, 15.02.2000, p. 1530-1535.

Research output: Contribution to journalArticle

Chong, James P J ; Hayashi, Mariko K. ; Simon, Martha N. ; Xu, Rui Ming ; Stillman, Bruce. / A double-hexamer archaeal minichromosome maintenance protein is an ATP- dependent DNA helicase. In: Proceedings of the National Academy of Sciences of the United States of America. 2000 ; Vol. 97, No. 4. pp. 1530-1535.
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