Wang H, Nicholson P R, Stillman D J
Department of Cellular, Viral and Molecular Biology, University of Utah School of Medicine, Salt Lake City 84132.
Mol Cell Biol. 1990 Apr;10(4):1743-53. doi: 10.1128/mcb.10.4.1743-1753.1990.
A DNA-binding protein has been identified from extracts of the budding yeast Saccharomyces cerevisiae which binds to sites present in the promoter regions of a number of yeast genes transcribed by RNA polymerase II, including SIN3 (also known as SDI1), SWI5, CDC9, and TOP1. This protein also binds to a site present in the enhancer for the 35S rRNA gene, which is transcribed by RNA polymerase I, and appears to be identical to the previously described REB1 protein (B. E. Morrow, S. P. Johnson, and J. R. Warner, J. Biol. Chem. 264:9061-9068, 1989). When oligonucleotides containing a REB1-binding site are placed between the CYC1 upstream activating sequence and TATA box, transcription by RNA polymerase II in vivo is substantially reduced, suggesting that REB1 acts as a repressor of RNA polymerase II transcription. The in vitro levels of the REB1 DNA-binding activity are reduced in extracts prepared from strains bearing a mutation in the SIN3 gene. A greater reduction in REB1 activity is observed if the sin3 mutant strain is grown in media containing galactose as a carbon source.
已从出芽酵母酿酒酵母的提取物中鉴定出一种DNA结合蛋白,它能与多种由RNA聚合酶II转录的酵母基因启动子区域中的位点结合,包括SIN3(也称为SDI1)、SWI5、CDC9和TOP1。这种蛋白还能与由RNA聚合酶I转录的35S rRNA基因增强子中的一个位点结合,并且似乎与先前描述的REB1蛋白相同(B.E.莫罗、S.P.约翰逊和J.R.华纳,《生物化学杂志》264:9061 - 9068,1989年)。当含有REB1结合位点的寡核苷酸置于CYC1上游激活序列和TATA框之间时,体内RNA聚合酶II的转录会大幅减少,这表明REB1作为RNA聚合酶II转录的阻遏物发挥作用。在携带SIN3基因突变的菌株制备的提取物中,REB1 DNA结合活性的体外水平降低。如果sin3突变菌株在以半乳糖作为碳源的培养基中生长,会观察到REB1活性有更大程度的降低。