In vivo interactions of archaeal Cdc6/Orc1 and minichromosome maintenance proteins with the replication origin.
Although genome analyses have suggested parallels between archaeal and eukaryotic replication systems, little is known about the DNA replication mechanism in Archaea. By two-dimensional gel electrophoreses we positioned a replication origin (oriC) within 1 kb in the chromosomal DNA of Pyrococcus abyssi, an anaerobic hyperthermophile, and demonstrated that the oriC is physically linked to the cdc6 gene. Our chromatin immunoprecipitation assays indicated that P. abyssi Cdc6 and minichromosome maintenance (MCM) proteins bind preferentially to the oriC region in the exponentially growing cells. Whereas the oriC association of MCM was specifically inhibited by stopping DNA replication with puromycin treatment, Cdc6 protein stayed bound to the replication origin after de novo protein synthesis was inhibited. Our data suggest that archaeal and eukaryotic Cdc6 and MCM proteins function similarly in replication initiation and imply that an oriC association of MCM could be regulated by an unknown mechanism in Archaea.
Matsunaga
F
F
Institut de Génétique et Microbiologie, Université de Paris-Sud, 91405 Orsay, France.
Forterre
P
P
Ishino
Y
Y
Myllykallio
H
H
D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D002874:Chromosome Mapping; D019847:Chromosomes, Archaeal / Q000235:genetics*; D016384:Consensus Sequence; D004261:DNA Replication; D019641:DNA, Archaeal / Q000737:chemistry / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006358:Hot Temperature; D008969:Molecular Sequence Data; D051738:Origin Recognition Complex; D019711:Pyrococcus / Q000235:genetics* / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D018741:Replication Origin; D029701:Saccharomyces cerevisiae Proteins
C066503:CDC6 protein, S cerevisiae; D018797:Cell Cycle Proteins; D002843:Chromatin; D019641:DNA, Archaeal; D004268:DNA-Binding Proteins; D051738:Origin Recognition Complex; D029701:Saccharomyces cerevisiae Proteins