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镁离子和锰离子对聚核糖胞苷酸及其2'-O-甲基衍生物的结构和逆转录酶模板活性的影响。

The effect of magnesium and manganese ions on the structure and template activity for reverse transcriptase of polyribocytidylate and its 2'-0-methyl derivative.

作者信息

Vamvakopoulos N C, Vournakis J N, Marcus S L

出版信息

Nucleic Acids Res. 1977 Oct;4(10):3589-97. doi: 10.1093/nar/4.10.3589.

DOI:10.1093/nar/4.10.3589
PMID:73165
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC342675/
Abstract

The secondary structure of the hydrogen bonded hybrids polycytidylate-oligodeoxguanylate (poly(rC)-(dG)12-18 and poly (2'-oMe) cytidylate-oligodeoxyguanylate (poly (rCm)-(dG)12-18 was studied at several magnesium and manganese ion concentrations. These hybrids are effective template-primer complexes for the synthesis of poly(dG) by avian myeloblastosis virus (AMV) DNA polymerase under disparate ionic conditions. Circular dichroism spectra and thermal melting data were obtained as a function of ion concentration, including conditions that allow optimum rates of poly (dG) synthesis by each complex. These studies demonstrate that both hybrids can change conformation and stability depending on their ionic environment. Comparison of enzyme activity and physical data suggest that the polymerase recognizes particular secondary structure features. Changes in the activity of the AMV polymerase can be induced by varying the Mg++ and Mn++ concentrations alone and in combination. These variations in enzyme activity are correlated with observed changes in the base-stacking alignment of the synthetic template primers. The ions, therefore, seem to affect enzyme activity by altering the conformation of the polnucleotide complexes.

摘要

在几种镁离子和锰离子浓度下，对氢键结合的杂合体聚胞苷酸 - 寡聚脱氧鸟苷酸（聚（rC） - （dG）12 - 18）和聚（2'-O-甲基）胞苷酸 - 寡聚脱氧鸟苷酸（聚（rCm） - （dG）12 - 18）的二级结构进行了研究。在不同的离子条件下，这些杂合体是禽成髓细胞瘤病毒（AMV）DNA聚合酶合成聚（dG）的有效模板 - 引物复合物。获得了作为离子浓度函数的圆二色光谱和热解链数据，包括允许每种复合物以最佳速率合成聚（dG）的条件。这些研究表明，两种杂合体都可以根据其离子环境改变构象和稳定性。酶活性和物理数据的比较表明，聚合酶识别特定的二级结构特征。单独或组合改变Mg++和Mn++浓度可诱导AMV聚合酶活性的变化。这些酶活性的变化与合成模板引物的碱基堆积排列中观察到的变化相关。因此，离子似乎通过改变多核苷酸复合物的构象来影响酶活性。

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Biochem Biophys Res Commun. 1969 Dec 4;37(6):895-901. doi: 10.1016/0006-291x(69)90215-0.
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