Schaefer T S, Sanders L K, Nathans D
Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205-2185, USA.
Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9097-101. doi: 10.1073/pnas.92.20.9097.
To identify proteins that regulate the transcriptional activity of c-Jun, we have used the yeast two-hybrid screen to detect mammalian polypeptides that might interact functionally with the N-terminal segment of c-Jun, a known regulatory region. Among the proteins identified is a short form of Stat3 (designated Stat3 beta). Stat3 beta is missing the 55 C-terminal amino acid residues of the long form (Stat3 alpha) and has 7 additional amino acid residues at its C terminus. In the absence of added cytokines, expression of Stat3 beta (but not Stat3 alpha) in transfected cells activated a promoter containing the interleukin 6 responsive element of the rat alpha 2-macroglobulin gene; coexpression of Stat3 beta and c-Jun led to enhanced cooperative activation of the promoter. Nuclear extracts of cells transfected with a Stat3 beta expression plasmid formed a complex with an oligonucleotide containing a Stat3 binding site, whereas extracts of cells transfected with a Stat3 alpha plasmid did not. We conclude that there is a short form of Stat3 (Stat3 beta), that Stat3 beta is transcriptionally active under conditions where Stat3 alpha is not, and that Stat3 beta and c-Jun are capable of cooperative activation of certain promoters.
为了鉴定调控c-Jun转录活性的蛋白质,我们利用酵母双杂交筛选来检测可能与c-Jun的N端片段(一个已知的调控区域)发生功能相互作用的哺乳动物多肽。鉴定出的蛋白质中有一个Stat3的短形式(命名为Stat3β)。Stat3β缺少长形式(Stat3α)的55个C端氨基酸残基,并且在其C端有另外7个氨基酸残基。在未添加细胞因子的情况下,转染细胞中Stat3β(而非Stat3α)的表达激活了一个含有大鼠α2-巨球蛋白基因白细胞介素6反应元件的启动子;Stat3β和c-Jun的共表达导致启动子的协同激活增强。用Stat3β表达质粒转染的细胞的核提取物与一个含有Stat3结合位点的寡核苷酸形成复合物,而用Stat3α质粒转染的细胞的提取物则不能。我们得出结论,存在Stat3的短形式(Stat3β),Stat3β在Stat3α不具有转录活性的条件下具有转录活性,并且Stat3β和c-Jun能够协同激活某些启动子。