Nomura T, Fujita N, Ishihama A
Nucleic Acids Res. 1986 Sep 11;14(17):6857-70. doi: 10.1093/nar/14.17.6857.
An in vitro mixed transcription system was employed to examine the possible alteration of the promoter selectivity of Escherichia coli RNA polymerase by specific tRNAs. Transcription in vitro was inhibited by most of the tRNAs examined, although the extent of the inhibition differed with the tRNA species. The inhibition by tRNAs was due to competition with DNA for binding RNA polymerase. This inhibitory effect remained after charging of the tRNAs with amino acids. The charging of tRNAfMet with fMet, but not with Met, abolished its inhibitory effect, and instead gave a stimulatory effect on the transcription from some promoters. These observations suggest that fMet-tRNAfMet plays a specific regulatory role in the coupling of transcription to translation.
采用体外混合转录系统来检测特定tRNA对大肠杆菌RNA聚合酶启动子选择性的可能改变。尽管不同种类的tRNA抑制程度有所不同,但大多数被检测的tRNA都会抑制体外转录。tRNA的抑制作用是由于与DNA竞争结合RNA聚合酶所致。给tRNA带上氨基酸后,这种抑制作用仍然存在。用甲硫氨酸而非甲硫氨酸对起始tRNA进行氨酰化修饰,消除了其抑制作用,反而对某些启动子的转录产生了刺激作用。这些观察结果表明,甲硫氨酰 - 起始tRNA在转录与翻译的偶联中发挥着特定的调节作用。