Park H, Francesconi S, Wang T S
Department of Pathology, Stanford University School of Medicine, California 94305-5324.
Mol Biol Cell. 1993 Feb;4(2):145-57. doi: 10.1091/mbc.4.2.145.
We have investigated the expression of two Schizosaccharomyces pombe replicative DNA polymerases alpha and delta during the cell cycle. The pol alpha+ and pol delta+ genes encoding DNA polymerases alpha and delta were isolated from S. pombe. Both pol alpha+ and pol delta+ genes are single copy genes in haploid cells and are essential for cell viability. In contrast to Saccharomyces cerevisiae homologs, the steady-state transcripts of both S. pombe pol alpha+ and pol delta+ genes were present throughout the cell cycle. Sequence analysis of the pol alpha+ and pol delta+ genes did not reveal the Mlu I motifs in their upstream sequences that are involved in cell cycle-dependent transcription of S. cerevisiae DNA synthesis genes as well as the S. pombe cdc22+ gene at the G1/S boundary. However, five near-match Mlu I motifs were found in the upstream region of the pol alpha+ gene. S. pombe DNA polymerases alpha and delta proteins were also expressed constantly throughout the cell cycle. In addition, the enzymatic activity of the S. pombe DNA polymerase alpha measured by in vitro assay was detected at all stages of the cell cycle. Thus, these S. pombe replicative DNA polymerases, like that of S. pombe cdc17+ gene, are expressed throughout the cell cycle at the transcriptional and protein level. These results indicate that S. pombe has at least two regulatory modes for the expression of genes involved in DNA replication and DNA precursor synthesis.
我们研究了粟酒裂殖酵母复制性DNA聚合酶α和δ在细胞周期中的表达情况。从粟酒裂殖酵母中分离出编码DNA聚合酶α和δ的pol α⁺和pol δ⁺基因。pol α⁺和pol δ⁺基因在单倍体细胞中均为单拷贝基因,对细胞活力至关重要。与酿酒酵母的同源物不同,粟酒裂殖酵母pol α⁺和pol δ⁺基因的稳态转录本在整个细胞周期中均存在。对pol α⁺和pol δ⁺基因的序列分析未在其上游序列中发现参与酿酒酵母DNA合成基因以及粟酒裂殖酵母cdc22⁺基因在G1/S边界的细胞周期依赖性转录的Mlu I基序。然而,在pol α⁺基因的上游区域发现了五个近似匹配的Mlu I基序。粟酒裂殖酵母DNA聚合酶α和δ蛋白在整个细胞周期中也持续表达。此外,通过体外测定法检测到的粟酒裂殖酵母DNA聚合酶α的酶活性在细胞周期的所有阶段均能检测到。因此,这些粟酒裂殖酵母复制性DNA聚合酶,就像粟酒裂殖酵母cdc17⁺基因一样,在转录和蛋白质水平上在整个细胞周期中均有表达。这些结果表明,粟酒裂殖酵母对于参与DNA复制和DNA前体合成的基因表达至少有两种调控模式。