Knaus R, Pollock R, Guarente L
Department of Biology, MIT, Cambridge 02139, USA.
EMBO J. 1996 Apr 15;15(8):1933-40.
Activation of transcription in eukaryotes depends upon the interplay between transcriptional activators and general transcription factors. While direct contacts between activators and general factors have been demonstrated in vitro, an additional class of proteins, termed co-activators, is also required of transcriptional activation. Here we describe a yeast protein, SUB1, that was isolated as a suppressor of the cold-sensitive TFIIB R78H mutant. The N-terminal third of SUB1 is highly similar to the mammalian co-activator PC4. We show that increased expression of SUB1 suppresses two alleles of TFIIB (E62G, R78H) specifically and that the deletion of SUB1 is lethal in combination with these same two alleles. We show that SUB1 binds to TFIIB in vitro and that it specifically inhibits the formation of TBP-TFIIB-promoter complexes. Furthermore we show that increasing the copy number of SUB1 stimulates transcriptional activation in vivo. Based on our results and recent observations of others, we propose that SUB1 plays a role in the release of TFIIB from the transcription complex during transcription initiation.
真核生物中,转录激活依赖于转录激活因子与通用转录因子之间的相互作用。虽然在体外已证实激活因子与通用因子之间存在直接接触,但转录激活还需要另一类蛋白质,即共激活因子。在此,我们描述了一种酵母蛋白SUB1,它是作为冷敏感型TFIIB R78H突变体的抑制子被分离出来的。SUB1的N端三分之一与哺乳动物共激活因子PC4高度相似。我们发现,SUB1表达量增加可特异性抑制TFIIB的两个等位基因(E62G、R78H),并且SUB1缺失与这两个相同等位基因结合时是致死的。我们表明,SUB1在体外与TFIIB结合,并且它特异性抑制TBP - TFIIB - 启动子复合物的形成。此外,我们还表明,增加SUB1的拷贝数可在体内刺激转录激活。基于我们的研究结果以及其他人最近的观察,我们提出SUB1在转录起始过程中从转录复合物释放TFIIB的过程中发挥作用。