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DNA polymerase ε encoded by cdc20 + is required for chromosomal DNA replication in the fission yeast Schizosaccharomyces pombe
Author(s) -
Sugino Akio,
Ohara Takeshi,
Sebastian Josef,
Nakashima Naomi,
Araki Hiroyuki
Publication year - 1998
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1046/j.1365-2443.1998.00169.x
Subject(s) - schizosaccharomyces pombe , biology , dna replication , schizosaccharomyces , control of chromosome duplication , fission , s phase , yeast , genetics , microbiology and biotechnology , dna , eukaryotic dna replication , saccharomyces cerevisiae , physics , quantum mechanics , neutron
Background: DNA polymerase II (PolII), the homologue of mammalian DNA polymerase ε, is essential for chromosomal DNA replication in the budding yeast Saccharomyces cerevisiae and also participates in S‐phase checkpoint control. An important issue is whether chromosomal DNA replication in other eukaryotes, including the fission yeast Schizosaccharomyces pombe —in which the characteristics of replication origins are poorly defined—also requires DNA polymerase ε. It has been shown that DNA polymerase ε is not required for the in vitro replication of SV40 DNA by human cell extracts. Results: We have cloned and sequenced S. pombe pol2 + , which is identical to the cell‐cycle gene cdc20 + , encoding the catalytic polypeptide of DNA polymerase ε (Polε). The predicted amino acid sequence of Polε is highly homologous to that of S. cerevisiae PolII and human Polε. Consistent with this, the Polε polypeptide was recognized by polyclonal antibodies against S. cerevisiae PolII holoenzyme (PolII*). The terminal morphology of cells containing the disrupted pol2 gene was similar to that of DNA replication mutant cells and cdc20 mutant cells. Furthermore, the Polε activity from temperature‐sensitive S. pombe cdc20 mutant cells was temperture‐sensitive, and chromosomal DNA replication in the mutant cells was inhibited at the restrictive temperatures. Conclusion: These data strongly suggest that Polε is required for normal chromosomal DNA replication in S. pombe , as is PolII in S. cerevisiae . Thus, eukaryotic chromosomal DNA is replicated differently from that of viral SV40 DNA.