Laimins L A, Khoury G, Gorman C, Howard B, Gruss P
Proc Natl Acad Sci U S A. 1982 Nov;79(21):6453-7. doi: 10.1073/pnas.79.21.6453.
The simian virus (SV40) 72-base pair (bp) tandem repeated sequences have recently been shown to function as activators or enhancers of early viral transcription. A recombinant viral genome was recently constructed by inserting 72-bp tandem repeats from the Moloney murine sarcoma virus (MSV) in place of the 72-bp repeats of SV40. Although this genome replicates in monkey kidney cells, its rate of large tumor antigen expression and replication is considerably slower than that of wild-type SV40. In mouse cells, however, equivalent levels of large tumor antigen appear to be expressed from both wild-type and recombinant genomes, suggesting a relationship between the level of enhancer activity and the host cell. To confirm this observation, we have applied a sensitive quantitative assay for gene expression based on the conversion of chloramphenicol to its acetylated forms. The gene encoding the enzymatic function chloramphenicol acetyltransferase was inserted into two vectors in which the enhancer sequences from SV40 or MSV were placed adjacent to the early SV40 promoter. The SV40 tandem repeats appear to activate gene expression to significantly higher levels in monkey kidney cells, but the MSV repeats are more active in two lines of mouse cells. These findings suggest that the tandem repeat elements may interact with host-specific molecules and, furthermore, may constitute one of the elements determining the host range of these eukaryotic viruses.
猿猴病毒(SV40)72碱基对(bp)串联重复序列最近已被证明可作为病毒早期转录的激活剂或增强子发挥作用。最近构建了一种重组病毒基因组,通过插入来自莫洛尼鼠肉瘤病毒(MSV)的72 bp串联重复序列来取代SV40的72 bp重复序列。尽管该基因组在猴肾细胞中复制,但其大肿瘤抗原表达和复制的速率比野生型SV40慢得多。然而,在小鼠细胞中,野生型和重组基因组似乎都表达了相当水平的大肿瘤抗原,这表明增强子活性水平与宿主细胞之间存在关联。为了证实这一观察结果,我们应用了一种基于氯霉素转化为其乙酰化形式的灵敏的基因表达定量测定法。将编码氯霉素乙酰转移酶酶功能的基因插入到两个载体中,其中来自SV40或MSV的增强子序列被置于SV40早期启动子附近。SV40串联重复序列似乎能在猴肾细胞中将基因表达激活到显著更高的水平,但MSV重复序列在两株小鼠细胞系中更具活性。这些发现表明,串联重复元件可能与宿主特异性分子相互作用,此外,可能构成决定这些真核病毒宿主范围的元件之一。