The RFC2 gene encoding a subunit of replication factor C of Saccharomyces cerevisiae

Vladimir Noskov, Satoko Maki, Yasuo Kawasaki, Sun hee Leem, Bun ichiro Ono, Hiroyuki Araki, Youri Pavlov, Akio Sugino

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

Replication Factor C (RF-C) of Saccharomyces cerevisiae is a complex that consists of several different polypeptides ranging from 120- to 37 kDa (Yoder and Burgers, 1991; Fien and Stillman, 1992), similar to human RF-C. We have isolated a gene, RFC2, that appears to be a component of the yeast RF-C. The RFC2 gene is located on chromosome X of S. cerevisiae and is essential for cell growth. Disruption of the RFC2 gene led to a dumbbell-shaped terminal morphology, common to mutants having a defect in chromosomal DNA replication. The steady-state levels of RFC2 mRNA fluctuated less during the cell cycle than other genes involved in DNA replication. Nucleotide sequence of the gene revealed an open reading frame corresponding to a polypeptide with a calculated Mr of 39,716 and a high degree of amino acid sequence homology to the 37-kDa subunit of human RF-C. Polyclonal antibodies against bacterially expressed Rfc2 protein specifically reduced RF-C activity in the RF-C-dependent reaction catalyzed by yeast DNA polymerase III. Furthermore, the Rfc2 protein was copurified with RF-C activity throughout RF-C purification. These results strongly suggest that the RFC2 gene product is a component of yeast RF-C. The bacterially expressed Rfc2 protein preferentially bound to primed single-strand DNA and weakly to ATP.

Original languageEnglish (US)
Pages (from-to)1527-1535
Number of pages9
JournalNucleic acids research
Volume22
Issue number9
DOIs
StatePublished - May 11 1994

ASJC Scopus subject areas

  • Genetics

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    Noskov, V., Maki, S., Kawasaki, Y., Leem, S. H., Ono, B. I., Araki, H., Pavlov, Y., & Sugino, A. (1994). The RFC2 gene encoding a subunit of replication factor C of Saccharomyces cerevisiae. Nucleic acids research, 22(9), 1527-1535. https://doi.org/10.1093/nar/22.9.1527