Chong, J. P. J., Hayashi, M. K., Simon, M. N., Xu, R. M., Stillman, B. (February 2000) 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, 97 (4). pp. 1530-1535. ISSN 0027-8424
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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 thermo-autotrophicum 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. thermoaototrophicum MCM protein. The purified protein forms a complex that has a molecular mass of approximate to 850 kDa, consistent with formation of a double hexamer, The protein has an ATP-independent DNA-binding activity, a DMA-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.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | REPLICATION LICENSING SYSTEM replication licensing system ORIGIN RECOGNITION COMPLEX origin recognition complex COMPLETE GENOME SEQUENCE complete genome sequence MCM PROTEINS MCM proteins VIRAL ORIGIN viral origin CELL-CYCLE cell cycle T-ANTIGEN T antigen T-antigen XENOPUS Xenopus YEAST yeast INITIATION initiation |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > helicase |
| CSHL Authors: | |
| Communities: | CSHL labs > Stillman lab |
| Highlight: | Stillman, Bruce W. |
| Depositing User: | CSHL Librarian |
| Date: | February 2000 |
| Date Deposited: | 07 Mar 2012 18:51 |
| Last Modified: | 10 Sep 2019 19:25 |
| PMCID: | PMC26469 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/24949 |
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