Shore D, Nasmyth K
MRC Laboratory of Molecular Biology, Cambridge, England.
Cell. 1987 Dec 4;51(5):721-32. doi: 10.1016/0092-8674(87)90095-x.
A DNA binding protein (RAP1, previously called SBF-E) has been shown to bind to putative regulatory sites at both yeast mating-type silencers, yet is not the product of genetically identified regulators of the silent loci. Here, we report the purification of RAP1 by DNA affinity chromatography, and the isolation of its gene from a lambda gt11 genomic library using antibodies raised against the protein. Disruption of the chromosomal copy of this gene is lethal. We show that RAP1 protein also binds in vitro to the upstream activation site (UAS) of MAT alpha and ribosomal protein genes. In addition, we show that two different UAS-associated RAP1 binding sites can substitute in vivo for a silencer binding site. Our results suggest that RAP1 may be a transcriptional regulator that can play a role in either repression or activation of transcription, depending upon the context of its binding site.
一种DNA结合蛋白（RAP1，以前称为SBF - E）已被证明能结合酵母交配型沉默子的假定调控位点，但它并非沉默位点经基因鉴定的调控因子的产物。在此，我们报告了通过DNA亲和层析对RAP1的纯化，以及使用针对该蛋白产生的抗体从λgt11基因组文库中分离其基因。该基因染色体拷贝的破坏是致死性的。我们表明，RAP1蛋白在体外也能结合MATα和核糖体蛋白基因的上游激活序列（UAS）。此外，我们还表明，两种不同的与UAS相关的RAP1结合位点在体内可替代沉默子结合位点。我们的结果表明，RAP1可能是一种转录调节因子，根据其结合位点的背景，它可以在转录的抑制或激活中发挥作用。
