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酿酒酵母CDC6基因在有丝分裂后期转录,编码一种控制S期起始的ATP/GTP酶。

The Saccharomyces cerevisiae CDC6 gene is transcribed at late mitosis and encodes a ATP/GTPase controlling S phase initiation.

作者信息

Zwerschke W, Rottjakob H W, Küntzel H

机构信息

Max-Planck-Institut für Experimentelle Medizin, Göttingen, Federal Republic of Germany.

出版信息

J Biol Chem. 1994 Sep 16;269(37):23351-6.

PMID:8083240
Abstract

We have compared the periodic fluctuation of mRNAs encoded by CDC6, a cell cycle gene controlling initiation of DNA replication, and CLN1, a G1 cyclin gene expressed at late G1. The maxima of CDC6 mRNA precede those of CLN1 mRNA by about 15 min in cells synchronized by release from pheromone arrest or from a cdc15 thermal arrest. CDC6 mRNA accumulates in cdc15-arrested telophase cells, decays around cell separation, and reappears during telophase and nuclear division of the next cycle. CDC6 transcription at late mitosis is not affected by the pheromone signaling pathway. The CDC6 mRNA fluctuation pattern is imposed to a CLN1-derived reporter gene if fused to the CDC6 promoter. The CDC6 gene was expressed in Escherichia coli as a glutathione-S-transferase fusion protein and purified by affinity chromatography. The Cdc6 protein binds rATP and rGTP upon UV cross-linking and catalyzes the DNA-independent hydrolysis of purine nucleoside triphosphates, but does not appear to interact directly with DNA. The Cdc6 protein may control the ATP-dependent initiation of replication by conferring ATPase activity to an origin-recognizing complex.

摘要

我们比较了由控制DNA复制起始的细胞周期基因CDC6和在G1晚期表达的G1细胞周期蛋白基因CLN1编码的mRNA的周期性波动。在通过从信息素阻滞或cdc15热阻滞释放而同步化的细胞中,CDC6 mRNA的最大值比CLN1 mRNA的最大值提前约15分钟。CDC6 mRNA在cdc15阻滞的末期细胞中积累,在细胞分离时衰减,并在下一个周期的末期和核分裂期间重新出现。有丝分裂后期的CDC6转录不受信息素信号通路的影响。如果与CDC6启动子融合,CDC6 mRNA的波动模式会强加于源自CLN1的报告基因。CDC6基因在大肠杆菌中作为谷胱甘肽-S-转移酶融合蛋白表达,并通过亲和层析纯化。紫外线交联后,Cdc6蛋白结合rATP和rGTP,并催化嘌呤核苷三磷酸的非DNA依赖性水解,但似乎不直接与DNA相互作用。Cdc6蛋白可能通过赋予识别起始位点的复合物ATPase活性来控制ATP依赖的复制起始。

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