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首页> 外文期刊>Molecular and Cellular Biology >Site-Specific DNA Binding of the Schizosaccharomyces pombe Origin Recognition Complex Is Determined by the Orc4 Subunit
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Site-Specific DNA Binding of the Schizosaccharomyces pombe Origin Recognition Complex Is Determined by the Orc4 Subunit

机译：Orc4亚基确定裂殖酵母起源识别复合物的特定于站点的DNA结合。

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The mechanism by which origin recognition complexes (ORCs) identify replication origins was investigated using purified Orc proteins from Schizosaccharomyces pombe. Orc4p alone bound tightly and specifically to several sites withinS. pombe replication origins that are genetically required for origin activity. These sites consisted of clusters of A or T residues on one strand but were devoid of either alternating A and T residues or GC-rich sequences. Addition of a complex consisting of Orc1, -2, -3, -5, and -6 proteins (ORC-5) altered neither Orc4p binding to origin DNA nor Orc4p protection of specific sequences. ORC-5 alone bound weakly and nonspecifically to DNA; strong binding required the presence of Orc4p. Under these conditions, all six subunits remained bound to chromatin isolated from each phase of the cell division cycle. These results reveal that the S.pombe ORC binds to multiple, specific sites within replication origins and that site selection, at least in vitro, is determined solely by the Orc4p subunit.
机译：使用来自 Schizosaccharomyces pombe 的纯化Orc蛋白，研究了起源识别复合物（ORC）识别复制起源的机制。 Orc4p单独紧密地且特异性地与 S 中的多个位点结合。起源活动在遗传上需要的 pombe 复制起源。这些位点由一条链上的A或T残基簇组成，但没有交替的A和T残基或富含GC的序列。添加由Orc1，-2，-3，-5和-6蛋白组成的复合物（ORC-5）既不会改变Orc4p与原始DNA的结合，也不会改变Orc4p对特定序列的保护。单独的ORC-5与DNA弱且非特异性结合;牢固的结合需要Orc4p的存在。在这些条件下，所有六个亚基仍与从细胞分裂周期各阶段分离的染色质结合。这些结果表明， S 。 pombe ORC与复制起点内的多个特定位点结合，并且至少在体外，该位点选择仅由Orc4p亚基决定。

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