碳二亚胺修饰对超螺旋PM2 DNA转录的抑制作用。
Inhibition of transcription of supercoiled PM2 DNA by carbodiimide modification.
作者信息
Flashner M S, Katopes M A, Lebowitz J
出版信息
Nucleic Acids Res. 1977 Jun;4(6):1713-26. doi: 10.1093/nar/4.6.1713.
Abstract
PM2 superhelican DNA (form I), which as been reacted with the single strand specific reagent, N-cyclohexyl-N'-beta-(methylmorpholinium)ethyl carbodiimide (CMC) is more than 95% inhibited in its ability to support transcription with E. coli B RNA polymerase in vitro. Almost complete inhibition of transcription was achieved after 2 hours of reaction with FI when only 1% of the bases were modified. A large increase in S20,* (from 26.8 S to 33.6 S) of FI DNA was observed during the course of reaction. Rifampicin resistant transcription is more susceptible to inhibition by CMC than total transcription, suggesting that the CMC is preferentially binding at promoter sites. These results clearly are in accord with the observation that supercoiled DNA contains localized regions of unpaired bases. The promotor sites for E. coli RNA polymerase in FI PM2 DNA appear to be located at or near these unpaired sites.
摘要
与单链特异性试剂N-环己基-N'-β-(甲基吗啉鎓)乙基碳二亚胺（CMC）反应后的PM2超螺旋DNA（I型），其在体外支持大肠杆菌B RNA聚合酶转录的能力受到超过95%的抑制。与FI反应2小时后，当只有1%的碱基被修饰时，转录几乎完全被抑制。在反应过程中观察到FI DNA的S20,*（从26.8 S增加到33.6 S）大幅增加。耐利福平转录比总转录更容易受到CMC的抑制，这表明CMC优先结合在启动子位点。这些结果显然与超螺旋DNA含有未配对碱基的局部区域这一观察结果一致。FI PM2 DNA中大肠杆菌RNA聚合酶的启动子位点似乎位于这些未配对位点处或其附近。
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