Gershowitz A, Moss B
J Virol. 1979 Sep;31(3):849-53. doi: 10.1128/JVI.31.3.849-853.1979.
Abortive transcription products were synthesized in vitro by UV-irradiated vaccinia virus particles that were incubated with all four ribonucleoside triphosphates or by unirradiated particles that were incubated in reaction mixtures deficient in CTP or UTP. The RNA sedimented at 4 to 6S in sucrose gradients, suggesting that premature termination had occurred, presumably in one case because the DNA contained UV-induced pyrimidine dimers and in the other case because of ribonucleoside triphosphate was present at limiting concentration for transcription. Nevertheless, the short transcripts were capped, methylated, and polyadenylylated, indicating that neither completion of an RNA chain nor processing from a polycistronic precursor was required for modification of either end of the RNA. In addition, the finding of m7G(5')pppAm and m7G(5')pppGm at the 5' ends of the short RNA molecules implied that transcription was initiated with both ATP and GTP. The presence of the polyadenylic acid tract suggested that a slow-down or cessation of transcription, rather than a specific 3'-terminal sequence, served as a signal for polyadenylylation.
将紫外线照射过的痘苗病毒颗粒与所有四种核糖核苷三磷酸一起孵育,或者将未照射的颗粒在缺乏CTP或UTP的反应混合物中孵育。RNA在蔗糖梯度中沉降在4至6S,这表明发生了过早终止,推测在一种情况下是因为DNA含有紫外线诱导的嘧啶二聚体,而在另一种情况下是因为核糖核苷三磷酸以限制转录的浓度存在。然而,短转录本被加帽、甲基化和聚腺苷酸化,这表明RNA链的完成或多顺反子前体的加工都不是RNA两端修饰所必需的。此外,在短RNA分子的5'端发现m7G(5')pppAm和m7G(5')pppGm意味着转录是由ATP和GTP共同起始的。聚腺苷酸序列的存在表明,转录的减慢或停止,而不是特定的3'-末端序列,作为聚腺苷酸化的信号。