ISOLATION OF THE GENE ENCODING THE SACCHAROMYCES-CEREVISIAE CENTROMERE-BINDING PROTEIN CP1
ISOLATION OF THE GENE ENCODING THE SACCHAROMYCES-CEREVISIAE CENTROMERE-BINDING PROTEIN CP1
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DOI：
10.1128/mcb.10.6.2458
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发表时间：
1990-06-01
影响因子：
5.3
通讯作者：
MASISON, DC
中科院分区：
文献类型：
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作者：
BAKER, RE;MASISON, DC
CP1 是酿酒酵母的序列特异性 DNA 结合蛋白，可识别酵母着丝粒高度保守的 DNA 元件 I (CDEI)。我们对编码 CPI 的基因进行了克隆和测序。该基因编码分子量为 39,400 的蛋白质。当在大肠杆菌中表达时，CP1基因指导CDEI结合蛋白的合成，其具有与纯化酵母CP1相同的凝胶迁移率。我们将 CP1 基因命名为 CEP1（着丝粒蛋白 1）。 CEP1 被定位并发现位于 X 染色体上，距 SUP4 2.0 厘摩。构建了大部分 CEP1 被删除的菌株。这些菌株缺乏可检测到的 CP1 活性并且具有活力；然而，CEP1 基因破坏导致细胞倍增时间增加 35%，有丝分裂染色体丢失率增加九倍。 CP1 基因破坏的一个意想不到的后果是与 cep1 遗传相关的蛋氨酸营养缺陷型。这一结果以及最近发现 MET25 启动子中的 CDE1 位点需要激活转录（D. Thomas、H. Cherest 和 Y. Surdin-Kerjan，J. Mol. Biol. 9：3292-3298，1989）表明 CP1 既是动粒蛋白又是转录因子。
CP1 is a sequence-specific DNA-binding protein of the yeast Saccharomyces cerevisiae which recognizes the highly conserved DNA element I (CDEI) of yeast centromeres. We cloned and sequenced the gene encoding CPI. The gene codes for a protein of molecular weight 39,400. When expressed in Escherichia coli, the CP1 gene directed the synthesis of a CDEI-binding protein having the same gel mobility as purified yeast CP1. We have given the CP1 gene the genetic designation CEP1 (centromere protein 1). CEP1 was mapped and found to reside on chromosome X, 2.0 centimorgans from SUP4. Strains were constructed in which most of CEP1 was deleted. Such strains lacked detectable CP1 activity and were viable; however, CEP1 gene disruption resulted in a 35% increase in cell doubling time and a ninefold increase in the rate of mitotic chromosome loss. An unexpected consequence of CP1 gene disruption was methionine auxotrophy genetically linked to cep1. This result and the recent finding that CDE1 sites in the MET25 promoter are required to activate transcription (D. Thomas, H. Cherest, and Y. Surdin-Kerjan, J. Mol. Biol. 9:3292-3298, 1989) suggest that CP1 is both a kinetochore protein and a transcription factor.