金精三羧酸对RNA指导的DNA聚合酶的抑制作用。
Inhibition of RNA-directed DNA polymerase by aurintricarboxylic acid.
作者信息
Givens J F, Manly K F
出版信息
Nucleic Acids Res. 1976 Feb;3(2):405-18. doi: 10.1093/nar/3.2.405.
Abstract
Commercial-grade aurintricarboxylic acid (ATA) inhibits poly(A), poly(C) and viral RNA-directed DNA synthesis by detergent-disrupted virions of Moloney murine leukemia virus. Paper chromatography of crude ATA yields two active components, which appear to behave identically, and at least two inactive components. The concentration of ATA needed to inhibit polymerase activity is proportional to the concentration of viral protein. The inhibition is neither attributable to contaminating heavy metal ions in the ATA preparation nor to chelation by ATA of Mn2+ or Zn2+, the necessary co-factors. Inhibition of the polymerase reaction by ATA greatly increases the Km for the primer [oligo(T)/oligo(dG)], while it only slightly lowers the Vmax and does not affect the Km's for the template [poly(A)/poly(C)] or the substrate (TTP/dGTP). Thus, ATA seems to reduce specifically the affinity of the polymerase for the DNA primer molecule.
摘要
商业级金精三羧酸（ATA）可抑制莫洛尼氏鼠白血病病毒经去污剂裂解的病毒粒子进行的聚腺苷酸、聚胞苷酸及病毒RNA指导的DNA合成。粗制ATA的纸层析产生两种活性成分，它们的行为似乎相同，以及至少两种无活性成分。抑制聚合酶活性所需的ATA浓度与病毒蛋白浓度成正比。这种抑制既不是由于ATA制剂中污染的重金属离子，也不是由于ATA对必需的辅助因子锰离子或锌离子的螯合作用。ATA对聚合酶反应的抑制极大地增加了对引物[寡聚（T）/寡聚（dG）]的米氏常数（Km），而它仅略微降低了最大反应速度（Vmax），并且不影响对模板[聚腺苷酸/聚胞苷酸]或底物（三磷酸胸苷/三磷酸鸟苷）的米氏常数。因此，ATA似乎特异性地降低了聚合酶对DNA引物分子的亲和力。
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