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首页> 外文期刊>Biochemistry >ATP cross-linked to Escherichia coli single-strand DNA-binding protein can be utilized by the catalytic center of primase as initiating nucleotide for primer RNA synthesis on phage G4oric template
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ATP cross-linked to Escherichia coli single-strand DNA-binding protein can be utilized by the catalytic center of primase as initiating nucleotide for primer RNA synthesis on phage G4oric template

机译：交联至大肠杆菌单链DNA结合蛋白的ATP可被引发酶的催化中心用作噬菌体G4oric模板上引物RNA合成的起始核苷酸

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摘要

We report a new observation of the role of Escherichia coli single-strand DNA binding protein (SSB) in synthesis of primer RNA (pRNA) catalyzed by E. coli primase on the SSB-coated phage G4oric template. Using a set of ATP priming substrates with reactive groups attached to the 5' gamma-phosphate on different length "arms", we have demonstrated that, in the primase/SSB/G4oric pRNA synthesis complex, ATP cross-linked to both primase and SSB could be equally utilized as initiating nucleotide for pRNA synthesis. The distance between SSB surface and ex-phosphorus of the priming substrate was estimated to be less than 7 Angstrom. ATP cross-linked to primase and SSB can be further elongated in the presence of other NTPs, giving almost identical patterns of covalently attached pRNAs of up to 12 nucleotides in length. The regions of primase and SSB with cross Linked ATP that can be used for pRNA synthesis are, therefore, arranged in a similar way relative to the active center of pRNA synthesis, The pRNA covalently linked to SSB was localized, mapping between Met48 and Trp88. This observation raises the possibility that SSB may play an active role in the initiation of pRNA synthesis in this system. [References: 23]
机译：我们报告了大肠杆菌单链DNA结合蛋白（SSB）在由SSB包被的噬菌体G4oric模板上被大肠杆菌引发酶催化的引物RNA（pRNA）合成中的作用的新观察。使用一组具有不同长度“臂”上连接到5'γ-磷酸的反应基团的ATP引发底物，我们证明了在primase / SSB / G4oric pRNA合成复合物中，ATP交联到primase和SSB上可以等同地用作pRNA合成的起始核苷酸。估计SSB表面与底涂底材的除磷之间的距离小于7埃。可以在存在其他NTP的情况下进一步延长与primase和SSB交联的ATP，从而获得长达12个核苷酸的共价连接的pRNA几乎相同的模式。因此，可用于pRNA合成的具有交联ATP的引发酶和SSB区域相对于pRNA合成的活性中心的排列方式相似。与SSB共价连接的pRNA被定位在Met48和Trp88之间。该观察结果提出了SSB在该系统中可能在pRNA合成起始中发挥积极作用的可能性。 [参考：23]

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