Abstract
Cdc6/Cdc18 is a conserved and essential component of prereplication complexes. The 2.0 A crystal structure of an archaeal Cdc6 ortholog, in conjunction with a mutational analysis of the homologous Cdc18 protein from Schizosaccharomyces pombe, reveals novel aspects of Cdc6/Cdc18 function. Two domains of Cdc6 form an AAA+-type nucleotide binding fold that is observed bound to Mg.ADP. A third domain adopts a winged-helix fold similar to known DNA binding modules. Sequence comparisons show that the winged-helix domain is conserved in Orc1, and mutagenesis data demonstrate that this region of Cdc6/Cdc18 is required for function in vivo. Additional mutational analyses suggest that nucleotide binding and/or hydrolysis by Cdc6/Cdc18 is required not only for progression through S phase, but also for maintenance of checkpoint control during S phase.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Alleles
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Cell Cycle Proteins* / chemistry
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Cell Cycle Proteins* / genetics
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Cell Cycle Proteins* / metabolism
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Cloning, Molecular
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Crystallography
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DNA Mutational Analysis
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DNA-Binding Proteins / genetics
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Fungal Proteins / chemistry
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Fungal Proteins / genetics
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Fungal Proteins / metabolism
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Gene Expression Regulation, Fungal
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Histones / chemistry
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Histones / genetics
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Histones / metabolism
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Molecular Sequence Data
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Mutation / physiology
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Nucleotides / metabolism
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Origin Recognition Complex
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Phenotype
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Protein Structure, Secondary
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Protein Structure, Tertiary
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Replication Origin / physiology*
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S Phase / physiology*
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Saccharomyces cerevisiae Proteins*
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Schizosaccharomyces
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Schizosaccharomyces pombe Proteins
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Sequence Homology, Amino Acid
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Structure-Activity Relationship
Substances
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CDC6 protein, S cerevisiae
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Cell Cycle Proteins
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DNA-Binding Proteins
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Fungal Proteins
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Histones
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Nucleotides
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Origin Recognition Complex
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Saccharomyces cerevisiae Proteins
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Schizosaccharomyces pombe Proteins
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cdc18 protein, S pombe