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

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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Noskov V,Maki S,Kawasaki Y,Leem SH,Ono B,Araki H,Pavlov Y,Sugino A

doi

10.1093/nar/22.9.1527

subject

Has Abstract

pub_date

1994-05-11 00:00:00

pages

1527-35

issue

9

eissn

0305-1048

issn

1362-4962

journal_volume

22

pub_type

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