NOVEL NON-TEMPLATED NUCLEOTIDE ADDITION-REACTIONS CATALYZED BY PROCARYOTIC AND EUKARYOTIC DNA-POLYMERASES
NOVEL NON-TEMPLATED NUCLEOTIDE ADDITION-REACTIONS CATALYZED BY PROCARYOTIC AND EUKARYOTIC DNA-POLYMERASES
复制标题
DOI：
10.1093/nar/16.20.9677
复制
发表时间：
1988-10-25
影响因子：
14.9
通讯作者：
CLARK, JM
中科院分区：
文献类型：
--
作者：
CLARK, JM
DNA聚合酶以模板导向的方式催化脱氧核糖核苷酸添加到DNA引物上。对模板指令的要求将这些酶与其他不使用模板的核苷酸转移酶（如末端脱氧核苷酸转移酶）区分开来。寡核苷酸底物被用来表征一种新的、非模板化的核苷酸加成反应，这种反应由来自多种原核生物和真核生物的DNA聚合酶进行。将脱氧核糖核苷酸添加到钝端DNA底物的3”羟基端，通过高分辨率变性聚丙烯酰胺凝胶电泳分析了反应产物。水热菌的DNA聚合酶，聚合酶。从鸡胚，大鼠聚合酶。禽成髓细胞病病毒逆转录酶和酿酒酵母DNA聚合酶均进行钝端加成反应。该反应需要双链DNA底物，但不需要模板链的编码信息。这些结果表明，模板指令不是DNA聚合酶催化核基转移反应的绝对要求。
DNA polymerases catalyze the addition of deoxyribonucleotides onto DNA primers in a template-directed manner. The requirement for template instruction distinguishes these enzymes from other nucleotidyl transferases, such as terminal deoxynucleotidyl transferase, that do not utilize a template. An oligonucleotide substrate was used to characterize a novel, non-templated nucleotide addition reaction carried out by DNA polymerases from a variety of procaryotic and eucaryotic sources. The products of the reaction, in which a deoxyribonucleotide was added to the 3'' hydroxyl terminus of a blunt-ended DNA substrate, were analyzed by electrophoresis on high resolution, denaturing polyacrylamide gels. DNA polymerase from Thermus aquaticus, polymerase .alpha. from chick embryo, rat polymerase .beta., reverse transcriptase from avian myeloblastosis virus, and DNA polymerase from Saccharomyces cerevisiae all carried out the blunt-end addition reaction. The reaction required a duplex DNA substrate but did not require coding information from the template strand. These results demonstrate that template instruction is not an absolute requirement for the catalysis of nucleotidy transfer reactions by DNA polymerases.