van Loon A E, Ligtenberg M, Reemst A M, Sussenbach J S, Rozijn T H
Laboratory for Physiological Chemistry, State University at Utrecht, The Netherlands.
Gene. 1987;58(1):109-26. doi: 10.1016/0378-1119(87)90034-5.
To determine whether differences in structure and organization of the transforming regions of Ad40 and Ad41 could explain the fastidious growth of these viruses, we have sequenced these regions and analyzed the structure of the corresponding mRNAs. These regions are 85% homologous to each other and exhibit 52% of homology to the analogous Ad5 region. Like the other adenoviruses, the Ad40 and Ad41 transforming regions contain two transcriptional units, E1a and E1b. Differences and similarities in the strategic sequences of these transcription units and Ad5 regulatory elements are discussed. Northern blotting and S1-nuclease analysis of RNA isolated from transformed cells reveal that in Ad40-transformed cells region E1a is transcribed into three partially overlapping mRNAs, while no transcription of region E1b can be detected. In Ad41-transformed cells only one E1a mRNA is found, comparable to the larger Ad40 E1a mRNA. In the Ad41 E1b region a single mRNA species is synthesized, which is comparable to the E1b messenger found in cells transformed by other adenovirus serotypes. Comparison of the large proteins encoded by the Ad40 and Ad41 E1a regions with the corresponding Ad5 E1a protein shows a relatively low homology in three conserved regions. These results are discussed in relation to the transforming capacity and the fastidious growth of Ad40 and Ad41 in conventional cell lines.
为了确定腺病毒40型(Ad40)和腺病毒41型(Ad41)转化区的结构和组织差异是否能够解释这两种病毒生长缓慢的特性,我们对这些区域进行了测序,并分析了相应mRNA的结构。这些区域彼此之间有85%的同源性,与腺病毒5型(Ad5)的类似区域有52%的同源性。与其他腺病毒一样,Ad40和Ad41的转化区包含两个转录单元,即E1a和E1b。本文讨论了这些转录单元的关键序列与Ad5调控元件之间的异同。对从转化细胞中分离出的RNA进行Northern印迹分析和S1核酸酶分析,结果显示,在Ad40转化的细胞中,E1a区域转录生成三种部分重叠的mRNA,而未检测到E1b区域的转录。在Ad41转化的细胞中,只发现一种E1a mRNA,与Ad40较大的E1a mRNA相当。在Ad41的E1b区域,合成了一种单一的mRNA,与其他腺病毒血清型转化细胞中发现的E1b信使RNA相当。将Ad40和Ad41的E1a区域编码的大蛋白与相应的Ad5 E1a蛋白进行比较,结果显示在三个保守区域中同源性相对较低。本文结合Ad40和Ad41在传统细胞系中的转化能力和生长缓慢的特性对这些结果进行了讨论。