Duesberg P, Helm K V, Canaani E
Proc Natl Acad Sci U S A. 1971 Oct;68(10):2505-9. doi: 10.1073/pnas.68.10.2505.
Several natural RNAs were compared with respect to their template activities for the DNA polymerase of Rous Sarcoma Virus during a 2-hr incubation period. 60-70S viral RNA was found to be a 5- to 10-fold better template than heat-dissociated Rous viral RNA, influenza virus RNA, tobacco mosaic virus RNA, or ribosomal RNA. Denatured salmon DNA is a little better, and poly(dAT) is 2-4 times better as a template for the enzyme than is 60-70S Rous viral RNA. The 60-70S RNAs of different strains of avian tumor viruses have very similar template activities for a given avian tumor virus DNA polymerase. Oligo(dT) or oligo(dC) were found to enhance the template activity of heat-dissociated Rous viral RNA 20- to 30-fold, and that of other natural RNAs tested one- to several-fold. DNA syntheses of 1-24% were obtained during a 2-hour incubation of the enzyme with the above RNA templates. The results suggest that the enzyme prefers partially doublestranded or hybrid regions of RNAs for optimal DNA synthesis, but certain regions of single-stranded RNA can also serve as templates.Poly(dAT) competes with viral RNA for purified DNA polymerase during DNA synthesis, as would be expected if RNA- and DNA-dependent DNA synthesis was performed by at least one common active site of the same enzyme.
在2小时的孵育期内,比较了几种天然RNA作为劳斯肉瘤病毒DNA聚合酶模板的活性。发现60 - 70S病毒RNA作为模板的效果比热解离的劳斯病毒RNA、流感病毒RNA、烟草花叶病毒RNA或核糖体RNA好5到10倍。变性鲑鱼DNA作为模板稍好一些,而聚(dAT)作为该酶的模板比60 - 70S劳斯病毒RNA好2至4倍。对于给定的禽肿瘤病毒DNA聚合酶,不同禽肿瘤病毒株的60 - 70S RNA具有非常相似的模板活性。发现寡聚(dT)或寡聚(dC)可将热解离的劳斯病毒RNA的模板活性提高20至30倍,将其他测试的天然RNA的模板活性提高1至数倍。在该酶与上述RNA模板孵育2小时期间,DNA合成率为1%至24%。结果表明,该酶进行最佳DNA合成时更喜欢RNA的部分双链或杂交区域,但单链RNA的某些区域也可作为模板。在DNA合成过程中,聚(dAT)与病毒RNA竞争纯化的DNA聚合酶,正如如果RNA依赖性和DNA依赖性DNA合成由同一酶的至少一个共同活性位点进行时所预期的那样。