Chinnadurai G, Rho H M, Horton R B, Green M
J Virol. 1976 Oct;20(1):255-63. doi: 10.1128/JVI.20.1.255-263.1976.
We have identified two mRNA species transcribed from the adenovirus 2 genome section (HindIII-G fragment) believed to harbor genes for initiation and maintenance of cell transformation. The HindIII-G fragment occupies the left 7.5% of the genome and is transcribed from left to right [poly(U:G) r strand]. Poly(A)-terminated labeled mRNA was isolated from polyribosomes of adenovirus 2 early infected KB cells and from the transformed cell line 8617, hybridization purified using the HindIII-G fragment, and electrophoresed on formamide-polyacrylamide gels. Viral mRNA's of 24S (1.2 X 10(6) daltons) and 14S (4.5 X 10(5) daltons) were isolated from early infected cells and of 22S (1.0 X 10(6) daltons) and 14S from 8617 cells. Hybridization competition indicated that HindIII-G-specific mRNA was present in the polysomes at one-sixth the concentration late after infection as compared with early, indicating that the proteins coded by the transforming segment may be synthesized at reduced amounts during late stages. Only 1/10 the amount of RNA labeled late annealed to the G fragment as compared with that labeled early (per weight of RNA). Thus, synthesis of transforming gene mRNA is probably "turned off" late after infection. Both 24S (22S) and 14S mRNA's from infected and 8617 cells were complementary to the Hpa I-E fragment (left 4.1% of genome). The Hpa I-E fragment is too small to encode 24S and 14S species, which implies that the 5'-terminal regions of both species are coded by the same DNA sequences.
我们已经鉴定出从腺病毒2基因组片段(HindIII - G片段)转录而来的两种mRNA,该片段被认为含有启动和维持细胞转化的基因。HindIII - G片段占据基因组左侧的7.5%,并从左至右转录[poly(U:G) r链]。从腺病毒2早期感染的KB细胞的多核糖体以及转化细胞系8617中分离出聚腺苷酸化末端标记的mRNA,使用HindIII - G片段进行杂交纯化,并在甲酰胺 - 聚丙烯酰胺凝胶上进行电泳。从早期感染细胞中分离出24S(1.2×10⁶道尔顿)和14S(4.5×10⁵道尔顿)的病毒mRNA,从8617细胞中分离出22S(1.0×10⁶道尔顿)和14S的病毒mRNA。杂交竞争表明,与早期相比,感染后期多核糖体中HindIII - G特异性mRNA的浓度为早期的六分之一,这表明转化区段编码的蛋白质在后期可能以减少的量合成。与早期标记的RNA相比(按RNA重量计算),后期标记的RNA与G片段退火的量仅为早期的1/10。因此,转化基因mRNA的合成在感染后期可能被“关闭”。来自感染细胞和8617细胞的24S(22S)和14S mRNA均与Hpa I - E片段(基因组左侧4.1%)互补。Hpa I - E片段太小,无法编码24S和14S种类,这意味着这两种种类的5'末端区域由相同的DNA序列编码。