DNA primase-DNA polymerase alpha from simian cells: sequence specificity of initiation sites on simian virus 40 DNA.
DNA primase-DNA polymerase alpha from simian cells: sequence specificity of initiation sites on simian virus 40 DNA.
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来自猿猴细胞的 DNA 引发酶-DNA 聚合酶 α：猿猴病毒 40 DNA 上起始位点的序列特异性。
DOI：
10.1128/mcb.5.5.1170
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发表时间：
1985
影响因子：
5.3
通讯作者：
M. DePamphilis
中科院分区：
文献类型：
--
作者：
M. Yamaguchi;E. Hendrickson;M. DePamphilis
使用猿病毒 40 DNA、噬菌体 M13 病毒粒子 DNA 和几种同聚物的独特单链区域作为模板，通过 CV-1 细胞 DNA 引物酶-DNA 聚合酶 α 合成 (p)ppRNA-DNA 链。完整的 RNA 引物专门用 RNA 加帽酶标记，通常长度为 6 至 8 个核糖核苷酸，但其长度范围为 1 至 9 个碱基。长度为 1 至 4 个核糖核苷酸的完整 RNA 引物的比例为 14% 至 73%，具体取决于所使用的模板。在相同核苷酸起始的引物之间以及在不同位点起始的引物之间，RNA 引物长度各不相同。因此，RNA 引物的大小取决于模板序列。通过在模板序列上绘制新生 RNA-DNA 链的 5' 末端、识别 5' 末端核糖核苷酸并对一个 RNA 引物进行部分测序来识别起始位点。在猿猴病毒 40 个 DNA 上总共鉴定出 56 个起始事件，平均每 16 个碱基 1 个。有些网站比其他网站更受欢迎。起始位点的共有序列由 3'-dCTTT 或 3'-dCCC 组成，中心位于 7 至 25 个富含嘧啶的残基内； RNA引物的5'端与dT或dC互补。高 ATP/GTP 比率促进 3'-dCTTT 位点 RNA 引物合成的起始，而低 ATP/GTP 比率促进 3'-dCCC 位点起始。类似地，聚脱氧胸苷酸和聚脱氧胞苷酸是唯一有效的均聚物模板。因此，模板序列和核糖核苷三磷酸浓度决定了DNA引物酶-DNA聚合酶α使用哪些起始位点。值得注意的是，体外选择的起始位点与猿猴病毒 40 DNA 体内复制期间选择的起始位点显着不同。
Unique single-stranded regions of simian virus 40 DNA, phage M13 virion DNA, and several homopolymers were used as templates for the synthesis of (p)ppRNA-DNA chains by CV-1 cell DNA primase-DNA polymerase alpha. Intact RNA primers, specifically labeled with an RNA capping enzyme, were typically 6 to 8 ribonucleotides long, although their lengths ranged from 1 to 9 bases. The fraction of intact RNA primers 1 to 4 ribonucleotides long was 14 to 73%, depending on the template used. RNA primer length varied among primers initiated at the same nucleotide, as well as with primers initiated at different sites. Thus, the size of an RNA primer depended on template sequence. Initiation sites were identified by mapping 5' ends of nascent RNA-DNA chains on the template sequence, identifying the 5'-terminal ribonucleotide, and partially sequencing one RNA primer. A total of 56 initiation events were identified on simian virus 40 DNA, an average of 1 every 16 bases. Some sites were preferred over others. A consensus sequence for initiation sites consisted of either 3'-dCTTT or 3'-dCCC centered within 7 to 25 pyrimidine-rich residues; the 5' ends of RNA primers were complementary to the dT or dC. High ATP/GTP ratios promoted initiation of RNA primer synthesis at 3'-dCTTT sites, whereas low ATP/GTP ratios promoted initiation at 3'-dCCC sites. Similarly, polydeoxythymidylic acid and polydeoxycytidylic acid were the only effective homopolymer templates. Thus, both template sequence and ribonucleoside triphosphate concentrations determine which initiation sites are used by DNA primase-DNA polymerase alpha. Remarkably, initiation sites selected in vitro were strikingly different from initiation sites selected during simian virus 40 DNA replication in vivo.
回文和非回文序列在体外和体内阻止 DNA 合成中的作用。
影响因子：
5.6
作者：
Weaver,DT;DePamphilis,ML
通讯作者：
DePamphilis,ML
猴病毒 40 起源区域的小鼠引物酶起始位点。
DOI：
10.1073/pnas.81.8.2342
发表时间：
1984
影响因子：
11.1
作者：
Tseng,BY;Ahlem,CN
通讯作者：
Ahlem,CN
DNA 聚合酶 α 辅因子 C1C2 充当引物识别蛋白。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Pritchard,CG;Weaver,DT;Baril,EF;DePamphilis,ML
通讯作者：
DePamphilis,ML