Jannun R, Chinnadurai G
Virus Res. 1987 Feb;7(1):33-48. doi: 10.1016/0168-1702(87)90056-6.
The functional relatedness of the transforming genes (E1a and E1b) of adenovirus type 9 (group D) which induces mammary tumors in rats and those of the non-tumorigenic adenoviruses, Ad2 and Ad5 (group C) was examined. Transfection of established rat embryo cells with a DNA segment containing the E1a and E1b regions of Ad9 resulted in efficient transformation; similar results have been shown for group A, B and C Ads. In contrast to Ads of group A, B and C, Ad9 DNA containing the E1 region or the entire viral genome was unable to transform primary baby rat kidney (BRK) cells. The functional relatedness of genes encoded within the E1 region was compared using a mutant complementation assay in which various group C mutants defective in the entire E1 region or in the E1a or E1b regions alone as well as mutants defective exclusively within the 19K or 58K T antigens coding regions of E1b were coinfected with wild type (wt) Ad9 and tested for group C mutant DNA replication, virus production, or expression of early and late genes. These studies have shown that a defect in the entire E1 region of Ad2 could only be complemented poorly by Ad9; our earlier studies have shown that coinfection with Ad12 (group A) or Ad7 (group B) resulted in efficient complementation (Brusca and Chinnadurai (1981) J. Virol. 39, 300-305). Further analysis indicated that a defect in the E1a region could be complemented by the group D E1a region. The level of E1a complementation as judged by mutant DNA replication and activation of expression of mutant early viral genes was about one-fourth to one-fifth the level in 293 cells that constitutively express Ad5 E1a and E1b regions. Our results indicate that a defect in the E1b 19K T antigen, which leads to degradation of intracellular DNA in infected cells, could be complemented by the group D protein. However, a defect in the E1b 58K T antigen could not be efficiently complemented by the group D protein. Coinfection of group C mutants defective in the 58K T antigen and Ad9 wt did not lead to an increase in the mutant viral production. Furthermore, in cells coinfected with the 58K T antigen mutants and Ad9 wt there was a large reduction in the accumulation of group C late cytoplasmic RNA. The observed complementation defect of Ad9 in supporting multiplication of group C mutants defective in the entire E1 region may therefore be a cumulative effect of both E1a and E1b regions.
对可在大鼠中诱发乳腺肿瘤的9型腺病毒（D组）的转化基因（E1a和E1b）与非致瘤性腺病毒Ad2和Ad5（C组）的转化基因之间的功能相关性进行了研究。用含有Ad9的E1a和E1b区域的DNA片段转染已建立的大鼠胚胎细胞，可实现高效转化；A组、B组和C组腺病毒也有类似结果。与A组、B组和C组腺病毒不同，含有E1区域或整个病毒基因组的Ad9 DNA无法转化原代新生大鼠肾（BRK）细胞。利用突变体互补分析比较了E1区域内编码基因的功能相关性，在该分析中，将整个E1区域或单独的E1a或E1b区域存在缺陷的各种C组突变体，以及仅在E1b的19K或58K T抗原编码区域内存在缺陷的突变体与野生型（wt）Ad9共感染，并检测C组突变体DNA复制、病毒产生或早期和晚期基因的表达情况。这些研究表明，Ad2的整个E1区域缺陷只能被Ad9低效互补；我们早期的研究表明，与Ad12（A组）或Ad7（B组）共感染可实现高效互补（布鲁斯卡和钦纳杜赖（1981年），《病毒学杂志》39，300 - 305）。进一步分析表明，E1a区域缺陷可被D组E1a区域互补。根据突变体DNA复制和突变体早期病毒基因表达激活判断，E1a互补水平约为组成型表达Ad5 E1a和E1b区域的293细胞中互补水平的四分之一到五分之一。我们的结果表明，导致受感染细胞内DNA降解的E1b 19K T抗原缺陷可被D组蛋白互补。然而E1b 58K T抗原缺陷不能被D组蛋白有效互补。58K T抗原缺陷的C组突变体与Ad9 wt共感染不会导致突变病毒产生增加。此外，在与58K T抗原突变体和Ad9 wt共感染的细胞中，C组晚期细胞质RNA的积累大幅减少。因此，观察到的Ad9在支持整个E1区域存在缺陷的C组突变体增殖方面的互补缺陷可能是E1a和E1b区域的累积效应。
