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

The minichromosome maintenance (MCM) proteins are essential for DNA replica tion 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 Methanobacteri um thermo-autotrophicum has allowed us to identify only a single MCM-like g ene (ORF Mt1770). This gene is most similar to MCM4 in eukaryotic cells. He re we have expressed and purified the M. thermoaototrophicum MCM protein. T he purified protein forms a complex that has a molecular mass of approximat e to 850 kDa, consistent with formation of a double hexamer, The protein ha s an ATP-independent DNA-binding activity, a DMA-stimulated ATPase activity that discriminates between single- and double-stranded DNA, and a strand-d isplacement (helicase) activity that can unwind up to 500 base pairs. The 3 ' to 5' helicase activity requires both ATP hydrolysis and a functional nuc leotide-binding site. Moreover, the double hexamer form is the active helic ase, It is therefore likely that an MCM complex acts as the replicative DNA helicase in eukaryotes and archaea, The simplified replication machinery i n archaea may provide a simplified model for assembly of the machinery requ ired for initiation of eukaryotic DNA replication.