Berk A J, Sharp P A
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274-8. doi: 10.1073/pnas.75.3.1274.
Biochemical methods are presented for determining the structure of spliced RNAs present in cells at low concentrations. Two cytoplasmic spliced viral RNAs were detected in CV-1 cells during the early phase of simian virus 40 (SV40) infection. One is 2200 nucleotides in length and is composed of two parts, 330 and 1900 nucleotides, mapping from approximately 0.67 to approximately 0.60 and from approximately 0.54 to approximately 0.14, respectively, on the standard viral map. The other is 2500 nucleotides long and also is composed of two parts, 630 and 1900 nucleotides mapping from approximately 0.67 to approximately 0.54 and from approximately 0.54 to approximately 0.14, respectively. Correlation of the structure of these mRNAs with the structure of the early SV40 proteins, small T antigen (17,000 daltons) and large T antigen (90,000 daltons), determined by others suggests that: (i) translation of the 2500-nucleotide mRNA yields small T antigen; (ii) translation of the 2200-nucleotide mRNA proceeds through the splice point in the RNA to produce large T antigen (and thus large T antigen is encoded in two separate regions of the viral genome); and (iii) the DNA sequences between approximately 0.67 and approximately 0.60 present in both mRNAs are translated in the same reading frame in both mRNAs to yield two separate gene products that have the same NH(2)-terminal sequence. Therefore, expression of the early SV40 genes is partially controlled at the level of splicing of RNAs.
本文介绍了用于确定细胞中低浓度存在的剪接RNA结构的生化方法。在猿猴病毒40(SV40)感染的早期阶段,在CV - 1细胞中检测到两种细胞质剪接病毒RNA。一种长度为2200个核苷酸,由两部分组成,分别为330和1900个核苷酸,在标准病毒图谱上分别从约0.67至约0.60以及从约0.54至约0.14定位。另一种长2500个核苷酸,也由两部分组成,分别为630和1900个核苷酸,分别从约0.67至约0.54以及从约0.54至约0.14定位。这些mRNA的结构与其他人确定的早期SV40蛋白、小T抗原(17,000道尔顿)和大T抗原(90,000道尔顿)的结构之间的相关性表明:(i)2500核苷酸mRNA的翻译产生小T抗原;(ii)2200核苷酸mRNA的翻译通过RNA中的剪接点进行,以产生大T抗原(因此大T抗原在病毒基因组的两个独立区域编码);(iii)两种mRNA中约0.67至约0.60之间的DNA序列在两种mRNA中以相同的阅读框进行翻译,以产生具有相同NH(2)-末端序列的两种独立基因产物。因此,早期SV40基因的表达在RNA剪接水平上受到部分控制。