Gray F C, MacNeill S A
Institute of Cell and Molecular Biology, University of Edinburgh, Scotland, UK.
Curr Genet. 2000 Mar;37(3):159-67. doi: 10.1007/s002940050514.
In the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe replication factor C (RF-C) plays key roles both in chromosomal DNA replication and in DNA replication checkpoint function. At the replication fork, the five-subunit RF-C complex functions to load the trimeric polymerase accessory factor PCNA onto DNA. PCNA then acts as a sliding clamp, tethering Pol delta to the DNA to maximise its processivity. Here we describe the cloning of the S. pombe rfc3+ gene, encoding a homologue of the S. cerevisiae Rfc3 and human hRFC36 proteins. The 1026 bp rfc3+ ORF is interrupted by five introns, ranging in size from 49 to 165 bp. The spliced ORF is predicted to encode a 342 amino-acid protein that is approximately 50% identical at the amino acid sequence level to the S. cerevisiae Rfc3 and human hRFC36 proteins. As expected, S. pombe rfc3+ is an essential gene, with rfc3delta cells being defective for DNA replication. Loss of rfc3+ function can be rescued by heterologous expression of either the S. cerevisiae Rfc3 or human hRFC36 proteins in S. pombe.
在酿酒酵母和粟酒裂殖酵母中,复制因子C(RF-C)在染色体DNA复制和DNA复制检查点功能中均发挥关键作用。在复制叉处,由五个亚基组成的RF-C复合物负责将三聚体聚合酶辅助因子PCNA加载到DNA上。然后,PCNA作为滑动夹,将Pol δ固定在DNA上以最大化其持续合成能力。在此,我们描述了粟酒裂殖酵母rfc3+基因的克隆,该基因编码酿酒酵母Rfc3和人类hRFC36蛋白的同源物。1026 bp的rfc3+开放阅读框被五个内含子打断,内含子大小从49到165 bp不等。剪接后的开放阅读框预计编码一个342个氨基酸的蛋白质,该蛋白质在氨基酸序列水平上与酿酒酵母Rfc3和人类hRFC36蛋白约50%相同。正如预期的那样,粟酒裂殖酵母rfc3+是一个必需基因,rfc3δ细胞在DNA复制方面存在缺陷。在粟酒裂殖酵母中,酿酒酵母Rfc3或人类hRFC36蛋白的异源表达可以挽救rfc3+功能的丧失。