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Multiple ORC‐binding sites are required for efficient MCM loading and origin firing in fission yeast
Author(s) -
Takahashi Tatsuro,
Ohara Eri,
Nishitani Hideo,
Masukata Hisao
Publication year - 2003
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/cdg079
Subject(s) - biology , schizosaccharomyces , minichromosome maintenance , origin of replication , origin recognition complex , chromatin , yeast , thymine , dna replication , dna , microbiology and biotechnology , biochemistry , schizosaccharomyces pombe , saccharomyces cerevisiae , eukaryotic dna replication
In most eukaryotes, replication origins are composed of long chromosome regions, and the exact sequences required for origin recognition complex (ORC) and minichromosome maintenance (MCM) complex association remain elusive. Here, we show that two stretches of adenine/thymine residues are collectively essential for a fission yeast chromosomal origin. Chromatin immunoprecipitation assays revealed that the ORC subunits are located within a 1 kb region of ori2004 . Analyses of deletion derivatives of ori2004 showed that adenine stretches are required for ORC binding in vivo . Synergistic interaction between ORC and adenine stretches was observed. On the other hand, MCM subunits were localized preferentially to a region near the initiation site, which is distant from adenine stretches. This association was dependent on adenine stretches and stimulated by a non‐adenine element. Our results suggest that association of multiple ORC molecules with a replication origin is required for efficient MCM loading and origin firing in fission yeast.