TEMPLATE-DIRECTED PAUSING IN INVITRO DNA-SYNTHESIS BY DNA POLYMERASE-ALPHA FROM DROSOPHILA-MELANOGASTER EMBRYOS
TEMPLATE-DIRECTED PAUSING IN INVITRO DNA-SYNTHESIS BY DNA POLYMERASE-ALPHA FROM DROSOPHILA-MELANOGASTER EMBRYOS
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DOI：
10.1073/pnas.79.4.983
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发表时间：
1982-01-01
期刊：
影响因子：
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通讯作者：
CLAYTON, DA
中科院分区：
文献类型：
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作者：
KAGUNI, LS;CLAYTON, DA
黑腹果蝇DNA聚合酶.α的活性。对 DNA 引发的单链 DNA 模板进行了检查。 DNA模板含有来自插入单链噬菌体载体M13Gori1的小鼠线粒体DNA重链起始区的1471个核苷酸序列。体外DNA合成暂停的优选位点被定位在克隆的mtDNA插入片段内和载体DNA内包含的G4c[互补]DNA链起点内。对复制中间体的新生 DNA 链的分析表明，暂停位点是离散的，既位于预测的稳定二联体的位置，又位于缺乏形成此类结构潜力的区域。观察到的大肠杆菌 DNA 聚合酶 III 全酶的动力学暂停位点模式在性质上与 DNA 聚合酶 α 中发现的相似。观察到的动力学暂停信号的子集在相似的条件下被大肠杆菌 DNA 聚合酶 I 识别。
The activity of D. melanogaster DNA polymerase .alpha. on DNA-primed single-stranded DNA templates was examined. The DNA templates contain a 1471-nucleotide sequence from the heavy-strand origin region of mouse mt[mitochondrial]DNA inserted into the single-stranded bacteriophage vector M13Gori1. Preferred sites for pausing of in vitro DNA synthesis were mapped within the cloned mtDNA insert and in the G4 c [complementary]DNA strand origin which is contained within the vector DNA. Analysis of nascent DNA strands from replicative intermediates has revealed that pause sites are discrete and lie both at the positions of predicted stable dyads and in regions lacking the potential for formation of such structures. The pattern of kinetic pause sites observed for Escherichia coli DNA polymerase III holoenzyme is qualitatively similar to that found for DNA polymerase .alpha.. A subset of the observed kinetic pause signals are recognized by E. coli DNA polymerase I under similar conditions.