TY - JOUR
T1 - The RFC2 gene encoding a subunit of replication factor C of Saccharomyces cerevisiae
AU - Noskov, Vladimir
AU - Maki, Satoko
AU - Kawasaki, Yasuo
AU - Leem, Sun hee
AU - Ono, Bun ichiro
AU - Araki, Hiroyuki
AU - Pavlov, Youri
AU - Sugino, Akio
N1 - Funding Information:
The RFC2 gene is essential for the mitotic growth, consistent with the idea that it is required for chromosomal DNA replication in yeast. This was supported by the terminal morphology (dumbbell) of cells in microcolonies raising from spores with the rfc2A mutation. Since mutant spores germinated and went through several rounds of the cell division before their death, there appear to be an excess of RF-C complex in the cell as is the case with yeast RF-A. Isolation of temperature-sensitive mutations will help to establish the role of RFC2 in chromosomal DNA replication.
PY - 1994/5/11
Y1 - 1994/5/11
N2 - 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.
AB - 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.
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U2 - 10.1093/nar/22.9.1527
DO - 10.1093/nar/22.9.1527
M3 - Article
C2 - 8202350
AN - SCOPUS:0028200657
SN - 0305-1048
VL - 22
SP - 1527
EP - 1535
JO - Nucleic acids research
JF - Nucleic acids research
IS - 9
ER -