ORC AND CDC6P INTERACT AND DETERMINE THE FREQUENCY OF INITIATION OF DNA-REPLICATION IN THE GENOME
ORC AND CDC6P INTERACT AND DETERMINE THE FREQUENCY OF INITIATION OF DNA-REPLICATION IN THE GENOME
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DOI：
10.1016/0092-8674(95)90528-6
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发表时间：
1995-06-02
期刊：
影响因子：
64.5
通讯作者：
STILLMAN, B
中科院分区：
文献类型：
--
作者：
LIANG, C;WEINREICH, M;STILLMAN, B
起源识别复合物（ORC）结合酵母S中的复制子。酿酒酵母中的一种，其方式与其作为DNA复制的起始蛋白一致。利用二维凝胶技术直接检测了温度敏感突变体orc的染色体DNA复制起始。与野生型细胞不同，在orc 2 -1和orc 5 -1突变细胞中，只有一部分复制子在允许温度下形成了DNA复制的活性起点。在限制温度下，活跃的复制因子的数量进一步减少。使用遗传筛选，CDC 6被鉴定为orc 5 -1的多拷贝抑制基因。二维凝胶电泳和生化分析表明Cdc 6p与ORC在功能和物理上相互作用。我们认为，ORC和Cdc 6p形成一个prereplication复合物在个别复制，因此合作，以确定在基因组中的DNA复制的起始频率。
The origin recognition complex (ORC) binds replicators in the yeast S. cerevisiae in a manner consistent with it being an initiator protein for DNA replication. Two-dimensional (2D) gel techniques were used to examine directly initiation of chromosomal DNA replication in temperature-sensitive orc mutants. Unlike in wild-type cells, in orc2-1 and orc5-1 mutant cells, only a subset of replicators formed active origins of DNA replication at the permissive temperature. At the restrictive temperature, the number of active replicators was diminished further. Using a genetic screen, CDC6 was identified as a multicopy suppressor of orc5-1. 2D gel and biochemical analyses demonstrated that Cdc6p interacted functionally and physically with ORC. We suggest that ORC and Cdc6p form a prereplication complex at individual replicators and therefore cooperate to determine the frequency of initiation of DNA replication in the genome.