Haegeman G, van Heuverswyn H, Gheysen D, Fiers W
J Virol. 1979 Aug;31(2):484-93. doi: 10.1128/JVI.31.2.484-493.1979.
d1-1811 is a viable simian virus 40 deletion mutant which lacks the DNA region corresponding to the major capping site of the late viral RNA. The exact size of the deletion (40 base pairs) was determined by comparison of the mutant DNA sequence with the wild-type simian virus 40 (strain 776) DNA sequence. Although d1-1811 forms somewhat smaller plaques, the amount of viral RNA late after infection was not significantly reduced compared with that of the wild type. Virus-specific, polyadenylate-containing, 32P-labeled late RNA was purified from the cytoplasm and enzymatically degraded to characterize the 5' terminus. The cap-containing oligonucleotides were isolated, and their structures were analyzed by further digestion. Instead of a single cap structure, we found a variety of capped 5' termini, with adenosine caps occurring much more frequently than guanosine caps. Nevertheless, there was a remarkable homology between both types of terminal sequences. Conceivably, the minor cap population present in wild-type simian virus 40 late mRNA may correspond to the collection of capped termini identified in the d1-1811 late mRNA . Cellular cytoplasmic RNA shows a similar pattern of cap structures, but the relative abundance is quite different.
d1 - 1811是一种可行的猿猴病毒40缺失突变体,它缺少与晚期病毒RNA主要加帽位点相对应的DNA区域。通过将突变体DNA序列与野生型猿猴病毒40(776株)DNA序列进行比较,确定了缺失的确切大小(40个碱基对)。尽管d1 - 1811形成的噬菌斑稍小,但感染后期病毒RNA的量与野生型相比并没有显著减少。从细胞质中纯化出病毒特异性的、含聚腺苷酸的、32P标记的晚期RNA,并进行酶促降解以表征其5'末端。分离出含帽寡核苷酸,并通过进一步消化分析其结构。我们发现,除了单一的帽结构外,还有多种加帽的5'末端,腺苷帽出现的频率比鸟苷帽高得多。然而,这两种末端序列之间存在显著的同源性。可以想象,野生型猿猴病毒40晚期mRNA中存在的少量帽结构可能与d1 - 1811晚期mRNA中鉴定出的加帽末端集合相对应。细胞细胞质RNA显示出类似的帽结构模式,但相对丰度有很大差异。