A multicopy suppressor gene of the Saccharomyces cerevisiae G1 cell cycle mutant gene dbf4 encodes a protein kinase and is identified as CDC5
A multicopy suppressor gene of the Saccharomyces cerevisiae G1 cell cycle mutant gene dbf4 encodes a protein kinase and is identified as CDC5
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酿酒酵母G1细胞周期突变基因dbf4的多拷贝抑制基因编码蛋白激酶，被鉴定为CDC5
DOI：
10.1128/mcb.13.7.4445-4457.1993
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发表时间：
1993
影响因子：
5.3
通讯作者：
Akio SUGINOl
中科院分区：
文献类型：
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作者：
K. Kitada;A. Johnson;L. Johnston;Akio SUGINOl
我们已经分离出一个多拷贝抑制剂的温度敏感的生长表型的生物体携带突变的DBF 4，一个基因，这是所需的启动染色体DNA复制在酿酒酵母和相互作用的CDC 7蛋白激酶。对暂时命名为MSD 2的抑制基因的核苷酸序列分析揭示了一个开放阅读框，其编码的蛋白质的计算M（r）为81，024，氨基酸序列与蛋白激酶的催化结构域相似。遗传连锁和互补分析表明，MSD 2是相同的细胞分裂周期基因CDC 5。磷酸化外源性添加的酪蛋白的活性通过抗TrpE-Cdc 5融合蛋白的抗血清免疫沉淀，所述TrpE-Cdc 5融合蛋白来自于在多拷贝质粒上含有CDC 5的野生型细胞的裂解物，但不是在质粒上的CDC 5的预测蛋白激酶结构域中具有小缺失的细胞的裂解物。CDC 5的缺失是致命的，并导致哑铃状的末端形态，细胞核几乎分裂但仍然相连。与在G2/M边界的功能一致，CDC 5转录物在细胞周期期间周期性地积累，在G2/M边界达到峰值。多拷贝质粒上的CDC 5还抑制温度敏感性cdc 15、cdc 20和dbf 2突变，这些突变影响细胞周期期间的有丝分裂。
We have isolated a multicopy suppressor of the temperature-sensitive growth phenotype of organisms carrying mutations of DBF4, a gene that is required for the initiation of chromosomal DNA replication in Saccharomyces cerevisiae and that interacts with the CDC7 protein kinase. Nucleotide sequence analysis of the suppressor gene, provisionally named MSD2, revealed an open reading frame encoding a protein with a calculated M(r) of 81,024, with amino acid sequence similarity to the catalytic domains of protein kinases. Both genetic linkage and complementation analyses indicated that MSD2 is identical to the cell division cycle gene CDC5. An activity that phosphorylated exogenously added casein was immunoprecipitated by antiserum against a TrpE-Cdc5 fusion protein from lysates of wild-type cells containing CDC5 on a multicopy plasmid but not of cells bearing a small deletion in the predicted protein kinase domain of CDC5 on the plasmid. Deletion of CDC5 was lethal and resulted in a dumbbell-shaped terminal morphology, with the nuclei almost divided but still connected. Consistent with the function at the G2/M boundary, the CDC5 transcript accumulated periodically during the cell cycle, peaking at the G2/M boundary. CDC5 on a multicopy plasmid also suppresses temperature-sensitive cdc15, cdc20, and dbf2 mutations which affect mitosis during the cell cycle.
用于构建trpE融合基因的具有多个克隆位点的高表达载体：pATH载体。
影响因子：
--
作者：
Koerner,TJ;Hill,JE;Myers,AM;Tzagoloff,A
通讯作者：
Tzagoloff,A
影响因子：
11.4
作者：
OSMANI, AH;ODONNELL, K;OSMANI, SA
通讯作者：
OSMANI, SA
来自酵母的 DNA 代谢基因 CDC7 编码丝氨酸（苏氨酸）蛋白激酶。
DOI：
10.1073/pnas.87.16.6272
发表时间：
1990
影响因子：
11.1
作者：
HollingsworthJr,RE;Sclafani,RA
通讯作者：
Sclafani,RA