The single minichromosome maintenance protein of Methanobacterium thermoautotrophicum DeltaH contains DNA helicase activity.

Previous studies have identified an ATP-dependent DNA helicase activity intrinsic to the human minichromosome maintenance (MCM) complex, composed of MCM subunits 4, 6, and 7 [Ishimi, Y. (1997) J. Biol. Chem. 272, 24508-24513]. In contrast to the presence of multiple MCM genes (at least six) in eukaryotes, the archaeon Methanobacterium thermoautotrophicum DeltaH (mth) genome contains a single open reading frame coding for an MCM protein. In this study we report the isolation of the mthMCM protein overexpressed in Escherichia coli. The purified recombinant protein was found to exist in both multimeric ( approximately 10(3) kDa) and monomeric (76 kDa) forms. Both forms of the protein bind to single-stranded DNA, hydrolyze ATP in the presence of DNA, and possess 3'-to-5' ATP-dependent DNA helicase activities. Thus, a single mthMCM protein contains biochemical properties identical to those associated with the eukaryotic MCM4, -6, and -7 complex. These results suggest that the characterization of the mthMCM protein and its multiple forms may contribute to our understanding of the role of MCM helicase activity in eukaryotic chromosomal DNA replication.

[1]  S. Pietrokovski,et al.  Isolation and Characterization of a Split B-type DNA Polymerase from the Archaeon Methanobacterium thermoautotrophicumΔH* , 1999, The Journal of Biological Chemistry.

[2]  Y. Kawarabayasi,et al.  Complete genome sequence of an aerobic hyper-thermophilic crenarchaeon, Aeropyrum pernix K1. , 1999, DNA research : an international journal for rapid publication of reports on genes and genomes.

[3]  B. Tye.  MCM proteins in DNA replication. , 1999, Annual review of biochemistry.

[4]  J. Leatherwood.  Emerging mechanisms of eukaryotic DNA replication initiation. , 1998, Current opinion in cell biology.

[5]  J. Diffley,et al.  Nucleotide-dependent prereplicative complex assembly by Cdc6p, a homolog of eukaryotic and prokaryotic clamp-loaders. , 1998, Molecular cell.

[6]  S. Forsburg,et al.  Multiple domains of fission yeast Cdc19p (MCM2) are required for its association with the core MCM complex. , 1998, Molecular biology of the cell.

[7]  S. Kearsey,et al.  MCM proteins: evolution, properties, and role in DNA replication. , 1998, Biochimica et biophysica acta.

[8]  B. Stillman,et al.  Formation of a preinitiation complex by S-phase cyclin CDK-dependent loading of Cdc45p onto chromatin. , 1998, Science.

[9]  H. Kimura,et al.  Biochemical Function of Mouse Minichromosome Maintenance 2 Protein* , 1998, The Journal of Biological Chemistry.

[10]  Tania A Baker,et al.  Polymerases and the Replisome: Machines within Machines , 1998, Cell.

[11]  F. Robb,et al.  Complete sequence and gene organization of the genome of a hyper-thermophilic archaebacterium, Pyrococcus horikoshii OT3. , 1998, DNA research : an international journal for rapid publication of reports on genes and genomes.

[12]  R. Fleischmann,et al.  The complete genome sequence of the hyperthermophilic, sulphate-reducing archaeon Archaeoglobus fulgidus , 1997, Nature.

[13]  G. Church,et al.  Complete genome sequence of Methanobacterium thermoautotrophicum deltaH: functional analysis and comparative genomics , 1997, Journal of bacteriology.

[14]  O. Aparicio,et al.  Components and Dynamics of DNA Replication Complexes in S. cerevisiae: Redistribution of MCM Proteins and Cdc45p during S Phase , 1997, Cell.

[15]  Y. Ishimi.  A DNA Helicase Activity Is Associated with an MCM4, -6, and -7 Protein Complex* , 1997, The Journal of Biological Chemistry.

[16]  K. Nasmyth,et al.  Loading of an Mcm Protein onto DNA Replication Origins Is Regulated by Cdc6p and CDKs , 1997, Cell.

[17]  M. Yanagida,et al.  A globular complex formation by Nda1 and the other five members of the MCM protein family in fission yeast , 1997, Genes to cells : devoted to molecular & cellular mechanisms.

[18]  L. Drury,et al.  Cdc6p-dependent loading of Mcm proteins onto pre-replicative chromatin in budding yeast. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[19]  S. Bell,et al.  Initiation of DNA replication in eukaryotic cells. , 1997, Annual review of cell and developmental biology.

[20]  H Kimura,et al.  Binding of Human Minichromosome Maintenance Proteins with Histone H3* , 1996, The Journal of Biological Chemistry.

[21]  I. Todorov,et al.  The role of MCM proteins in the cell cycle control of genome duplication , 1996, BioEssays : news and reviews in molecular, cellular and developmental biology.

[22]  J. Blow,et al.  The role of MCM/P1 proteins in the licensing of DNA replication. , 1996, Trends in biochemical sciences.

[23]  James P. J. Chong,et al.  Purification of an MCM-containing complex as a component of the DNA replication licensing system , 1995, Nature.

[24]  R. Knippers,et al.  Interactions of human nuclear proteins P1Mcm3 and P1Cdc46. , 1995, European journal of biochemistry.

[25]  F. Dean,et al.  The simian virus 40 T antigen double hexamer assembles around the DNA at the replication origin. , 1992, The Journal of biological chemistry.

[26]  A. Kwong,et al.  The in vitro replication of DNA containing the SV40 origin. , 1990, The Journal of biological chemistry.

[27]  H. Sambrook.  Molecular cloning : a laboratory manual. Cold Spring Harbor, NY , 1989 .

[28]  B. Tye,et al.  Mutants of S. cerevisiae defective in the maintenance of minichromosomes. , 1984, Genetics.

[29]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[30]  G. Weinstock,et al.  recA protein-catalyzed strand assimilation: stimulation by Escherichia coli single-stranded DNA-binding protein. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[31]  C. Woese,et al.  Phylogenetic structure of the prokaryotic domain: The primary kingdoms , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[32]  J. Zeikus,et al.  Methanobacterium thermoautotrophicus sp. n., an Anaerobic, Autotrophic, Extreme Thermophile , 1972, Journal of bacteriology.