Somasekhar M B, Mertz J E
Nucleic Acids Res. 1985 Aug 12;13(15):5591-609. doi: 10.1093/nar/13.15.5591.
The spliced species of late SV40 RNAs present in the cytoplasm of cells infected with various wild-type and mutant strains of SV40 that differ in their leader regions were determined using a novel modification of the primer extension method and the S1 nuclease mapping technique. These data indicated that mutations within the first exon of the late RNAs can affect dramatically the utilization of downstream donor and acceptor splice sites. In one instance, a ten base pair insertion within the predominant first exon increased utilization of an infrequently utilized donor splice site such that the small alteration became part of an intervening sequence, thereby suggesting a novel mechanism for regulation of gene expression. In addition, our method enabled detection of a previously unidentified spliced species, representing less than one percent of the SV40 late 19S RNA present in cells infected with wild-type virus, that may be an intermediate in the synthesis of a known doubly spliced 16S RNA species of SV40.
利用引物延伸法和S1核酸酶图谱技术的一种新改进,确定了感染不同野生型和突变型SV40菌株(其前导区不同)的细胞胞质中存在的晚期SV40 RNA的剪接种类。这些数据表明,晚期RNA第一个外显子内的突变可显著影响下游供体和受体剪接位点的利用。在一个实例中,主要第一个外显子内的一个10个碱基对的插入增加了一个很少使用的供体剪接位点的利用,使得这个小的改变成为一个居间序列的一部分,从而提示了一种新的基因表达调控机制。此外,我们的方法能够检测到一种以前未鉴定的剪接种类,它在感染野生型病毒的细胞中占SV40晚期19S RNA的不到1%,可能是SV40一种已知的双剪接16S RNA种类合成过程中的中间体。