DNA-dependent RNA polymerases from Acanthamoeba castellanii. Comparative subunit structures of the homogeneous enzymes.
DNA-dependent RNA polymerases from Acanthamoeba castellanii. Comparative subunit structures of the homogeneous enzymes.
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来自卡斯氏棘阿米巴的 DNA 依赖性 RNA 聚合酶。
DOI：
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发表时间：
1979
影响因子：
4.8
通讯作者：
M. Paule
中科院分区：
文献类型：
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作者：
J. D'Alessio;P. J. Perna;M. Paule
用几种电泳方法比较了卡氏阿米巴三种依赖DNA的RNA聚合酶的组成多肽。在十二烷基硫酸钠（SDS）存在下的聚丙烯酰胺凝胶电泳表明，许多多肽组分的同工酶具有相同的分子量。双向电泳（在8 M尿素中等电聚焦：SDS-聚丙烯酰胺凝胶电泳）表明，相同分子量的多肽也具有相同的等电pH值。这些多肽在酸性或碱性pH值下在8 M尿素中电泳后也是一致的，随后在SDS存在下进行第二次电泳分离。根据这些标准，分子量为13，300、15，500、17，500、22，500、37，000和39，000的亚基在聚合酶I和III中是不可区分的。13，300、15，500和22，500个亚基也由II类聚合酶共享。此外，在碱性条件下在8 M尿素中的电泳揭示了17，500分子量亚基的微观异质性。酵母和阿米巴之间的共同亚基的惊人相似的模式表明，一个通用的功能单位的安排可能是真核生物聚合酶的一个基本特征。
The constituent polypeptides of the three classes of DNA-dependent RNA polymerase from Acanthamoeba castellanii were compared by several electrophoretic methods. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) reveals that a number of polypeptide components of the isozymes have identical molecular weights. Two-dimensional electrophoresis (isoelectric focusing in 8 M urea:SDS-polyacrylamide gel electrophoresis) demonstrates that the polypeptides of identical molecular weights also have identical isoelectric pH values. These polypeptides were also coincident after electrophoresis in 8 M urea at acidic or basic pH values followed by a second electrophoretic separation in the presence of SDS. By these criteria, subunits of molecular weight 13,300, 15,500, 17,500, 22,500, 37,000, and 39,000 are indistinguishable in polymerase I and III. The 13,300, 15,500, and 22,500 subunits are also shared by the class II polymerase. In addition, electrophoresis in 8 M urea under basic conditions reveals microheterogeneity in the 17,500 molecular weight subunit. The strikingly similar pattern of common subunits between yeast and Acanthamoeba suggests that a universal arrangement of functional units may be an essential feature of the eukaryotic polymerases.