Laub O, Jakobovits E B, Aloni Y
Proc Natl Acad Sci U S A. 1980 Jun;77(6):3297-3301. doi: 10.1073/pnas.77.6.3297.
Short RNA chains initiating at the major promoter sites for simian virus 40 (SV40) late transcription are elongated to approximately 450 nucleotides in a molar ammount greater than that from any other region of the viral DNA. This conclusion is based on the following observations: (i) Transcriptional complexes isolated by Sarkosyl and by hypotonic leaching (minichromosomes) from nuclei of cells infected with SV40 as well as intact nuclei were pulse labeled in vitro with [alpha-32P]TUP and were observed to synthesize short RNA transcripts that hybridized predominantly to a SV40 DNA fragment spanning between 0.67 and 0.76 map units. (ii) In the presence of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), a drug known to accentuate premature transcriptional termination, accumulation of these short SV40 RNA chains was enhanced. When SV40-infected cells were pretreated with DRB and then labeled in vivo or in vitro, they synthesized short labeled viral RNAs that hydridized almost exclusively with the DNA fragment spanning between 0.67 and 0.76 map units. These observations suggest a mechanism in the regulation of SV40 late transcription.
起始于猴病毒40(SV40)晚期转录主要启动子位点的短RNA链,在摩尔量上会延伸至约450个核苷酸,且该摩尔量大于病毒DNA其他任何区域的摩尔量。这一结论基于以下观察结果:(i)用十二烷基肌氨酸钠和低渗浸出法(微型染色体)从感染SV40的细胞核以及完整细胞核中分离出的转录复合物,在体外用[α-32P]TUP进行脉冲标记,并观察到其合成的短RNA转录本主要与跨越0.67至0.76图距单位的SV40 DNA片段杂交。(ii)在5,6-二氯-1-β-D-呋喃核糖基苯并咪唑(DRB)存在的情况下,已知该药物会加剧转录提前终止,这些短SV40 RNA链的积累会增强。当用DRB预处理感染SV40的细胞,然后在体内或体外进行标记时,它们合成的短标记病毒RNA几乎只与跨越0.67至0.76图距单位的DNA片段杂交。这些观察结果提示了一种SV40晚期转录调控机制。