与小鼠胞质A颗粒相关的一种RNA指导的DNA聚合酶的特性分析。
Characterization of an RNA-directed DNA polymerase found in association with murine intracytoplasmic A-particles.
作者信息
Kohno M, Tanaka H
出版信息
J Virol. 1977 May;22(2):273-80. doi: 10.1128/JVI.22.2.273-280.1977.
Abstract
An RNA-directed DNA polymerase was found to be associated with intracytoplasmic A-particles from DBA/2 mouse leukemia cells. The enzyme activity was detected after disrupting the purified particles with 2 M NaCl-20 mM dithiothreitol. The presence of a divalent cation and all four deoxyribonucleoside triphosphates was essential for this enzyme activity. The enzyme had a clear preference for Mg2+ over Mn2+. Cesium sulfate isopycnic gradient centrifugation of the DNA product synthesized in the actinomycin D-containing reaction revealed the presence of DNA-RNA hybrid. Furthermore, the purified DNA product was found to hybridize with RNA isolated from A-particles. These observations strongly indicate that the endogenous A-particle RNA serves as the template for the DNA polymerase.
摘要
发现一种RNA指导的DNA聚合酶与来自DBA/2小鼠白血病细胞的胞质内A颗粒相关。在用2M NaCl-20mM二硫苏糖醇破坏纯化的颗粒后检测到该酶活性。二价阳离子和所有四种脱氧核糖核苷三磷酸的存在对该酶活性至关重要。该酶对Mg2+的偏好明显高于Mn2+。对在含放线菌素D的反应中合成的DNA产物进行硫酸铯等密度梯度离心，结果显示存在DNA-RNA杂交体。此外，发现纯化的DNA产物可与从A颗粒中分离的RNA杂交。这些观察结果有力地表明，内源性A颗粒RNA作为DNA聚合酶的模板。
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