Hoeffler W K, Kovelman R, Roeder R G
Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021.
Cell. 1988 Jun 17;53(6):907-20. doi: 10.1016/s0092-8674(88)90409-6.
The factor(s) responsible for the adenovirus E1A-stimulated transcription of RNA polymerase III genes was localized previously in a chromatographic fraction containing transcription factor IIIC (TFIIIC). In further studies, two distinct forms of TFIIIC, which were chromatographically separable, generated VA gene-protein complexes that were distinguished by gel shift assays. The form of TFIIIC that generated the more slowly migrating promoter complex had greater transcriptional activity in vitro, associated more rapidly with the promoter, and formed a more salt-resistant complex. Greater amounts of this more active form of TFIIIC resulted from either E1A expression during infection or growth of the cells in a higher concentration of serum, whereas template commitment assays indicated that overall TFIIIC concentrations remained unchanged during viral infection. The in vitro interconversion of the two forms of TFIIIC by phosphatase treatment suggests that transcriptional activation of RNA polymerase III genes can be mediated by phosphorylation of TFIIIC.
之前已将负责腺病毒E1A刺激的RNA聚合酶III基因转录的因子定位在含有转录因子IIIC(TFIIIC)的色谱级分中。在进一步的研究中,两种通过色谱可分离的不同形式的TFIIIC产生了VA基因-蛋白质复合物,通过凝胶迁移试验可区分这些复合物。产生迁移较慢的启动子复合物的TFIIIC形式在体外具有更高的转录活性,与启动子的结合更快,并形成更耐盐的复合物。感染期间的E1A表达或细胞在更高浓度血清中生长会产生更多这种活性更高的TFIIIC形式,而模板结合试验表明在病毒感染期间TFIIIC的总体浓度保持不变。通过磷酸酶处理使两种形式的TFIIIC在体外相互转化,这表明RNA聚合酶III基因的转录激活可由TFIIIC的磷酸化介导。