Kessler M, Aloni Y
Department of Molecular Genetics and Virology, Weizmann Institute of Science, Rehovot, Israel.
Gene. 1989 Dec 7;84(1):65-72. doi: 10.1016/0378-1119(89)90140-6.
We have previously reported that a mechanism resembling attenuation in prokaryotes regulates simian virus 40 (SV40) late gene expression. We have suggested that modulation of the attenuator RNA secondary structure is an integral element regulating the elongation block at the attenuation site [Hay et al., Cell 29 (1982) 183-193]. In the present study, oligodeoxyribonucleotides (oligos), 13-19 nucleotides long, were used to probe the involvement of the attenuator RNA secondary structure in the control of elongation block at the SV40 attenuation site. These oligos are complementary to segments of the attenuator RNA suggested to play a role in the regulation of attenuation. The oligos were added to an in vitro transcription reaction containing SV40 transcription complexes, and their effect on transcription through the attenuation site was measured. As predicted, the three oligos caused specific decreases in the elongation block at the SV40 attenuation site. These results provide direct evidence for the involvement of RNA secondary structure in the attenuation mechanism in SV40.
我们之前报道过，一种类似于原核生物衰减作用的机制调控着猴病毒40（SV40）晚期基因的表达。我们提出，衰减子RNA二级结构的调节是在衰减位点调控延伸阻断的一个不可或缺的要素[Hay等人，《细胞》29（1982）183 - 193]。在本研究中，长度为13 - 19个核苷酸的寡脱氧核糖核苷酸（oligos）被用于探究衰减子RNA二级结构在SV40衰减位点延伸阻断控制中的作用。这些oligos与衰减子RNA中被认为在衰减调控中起作用的片段互补。将这些oligos添加到含有SV40转录复合物的体外转录反应中，并测定它们对通过衰减位点转录的影响。正如所预测的，这三种oligos导致SV40衰减位点的延伸阻断特异性降低。这些结果为RNA二级结构参与SV40的衰减机制提供了直接证据。
