Ferguson B, Krippl B, Andrisani O, Jones N, Westphal H, Rosenberg M
Mol Cell Biol. 1985 Oct;5(10):2653-61. doi: 10.1128/mcb.5.10.2653-2661.1985.
We previously purified and characterized functionally the Escherichia coli-expressed product of the human subgroup C adenovirus E1A 13S mRNA (B. Ferguson, N. Jones, J. Richter, and M. Rosenberg, Science 224:1343-1346, 1984; B. Krippl, B. Ferguson, M. Rosenberg, and H. Westphal, Proc. Natl. Acad. Sci. USA 81:6988-6992, 1984). We have now expressed in E. coli and purified the protein product encoded by the human subgroup C adenovirus E1A 12S mRNA and have compared the functional properties of this protein with those of the E1A 13S mRNA product. Using microinjection techniques to introduce these proteins into mammalian cells, we found that the E1A 12S mRNA product, like the 13S mRNA product, localized rapidly to the cell nucleus and induced adenovirus gene expression. Although both E1A gene products localized to the nucleus and stimulated adenovirus gene transcription, these proteins did not directly bind to DNA under conditions in which a known DNA-binding protein, the human c-myc gene product, bound DNA efficiently. Thus, the E1A and myc gene products, which have been related both structurally and functionally, exhibit distinctly different biochemical properties.
我们之前对在大肠杆菌中表达的人C亚组腺病毒E1A 13S mRNA的产物进行了纯化和功能鉴定（B. Ferguson、N. Jones、J. Richter和M. Rosenberg，《科学》224:1343 - 1346，1984；B. Krippl、B. Ferguson、M. Rosenberg和H. Westphal，《美国国家科学院院刊》81:6988 - 6992，1984）。我们现在已在大肠杆菌中表达并纯化了人C亚组腺病毒E1A 12S mRNA编码的蛋白质产物，并将该蛋白质的功能特性与E1A 13S mRNA产物的功能特性进行了比较。利用显微注射技术将这些蛋白质导入哺乳动物细胞，我们发现E1A 12S mRNA产物与13S mRNA产物一样，能迅速定位于细胞核并诱导腺病毒基因表达。尽管两种E1A基因产物都定位于细胞核并刺激腺病毒基因转录，但在已知的DNA结合蛋白——人c - myc基因产物能有效结合DNA的条件下，这些蛋白质并不直接与DNA结合。因此，在结构和功能上都有关联的E1A和myc基因产物表现出明显不同的生化特性。
