Stillman D J, Bankier A T, Seddon A, Groenhout E G, Nasmyth K A
Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK.
EMBO J. 1988 Feb;7(2):485-94. doi: 10.1002/j.1460-2075.1988.tb02836.x.
The yeast HO gene, which encodes an endonuclease involved in initiating mating type interconversion, is expressed in mother cells but not in daughters. It has been demonstrated that the SWI5 gene, which is an activator of HO expression, plays a critical role in this differential mother/daughter expression of HO. In this paper we describe the cloning and sequencing of the SWI5 gene. The predicted amino acid sequence derived from the cloned SWI5 gene shows homology with the repeated DNA-binding domains ('zinc fingers') of Xenopus transcription factor TFIIIA. A region of the HO promoter involved in the SWI5-dependent transcriptional activation of HO was identified by deletion analysis of the HO promoter in the chromosome, and by testing the ability of HO DNA fragments to activate transcription in the context of a heterologous promoter. The SWI5 gene product was overproduced in yeast from the GAL1-10 promoter, since the SWI5 protein is made at very low levels in wild-type strains, and protein extracts were used to demonstrate that the SWI5 protein binds in vitro to a segment of the HO promoter required for transcriptional activation in vivo.
酵母HO基因编码一种参与启动交配型相互转换的内切核酸酶,该基因在母细胞中表达,而在子细胞中不表达。已证明,作为HO基因表达激活剂的SWI5基因,在HO基因这种母/子细胞差异表达中起关键作用。在本文中,我们描述了SWI5基因的克隆和测序。从克隆的SWI5基因推导出来的预测氨基酸序列,与非洲爪蟾转录因子TFIIIA的重复DNA结合结构域(“锌指”)具有同源性。通过对染色体上HO基因启动子进行缺失分析,以及通过检测HO基因DNA片段在异源启动子背景下激活转录的能力,确定了HO基因启动子中与SWI5依赖的HO基因转录激活有关的区域。由于SWI5蛋白在野生型菌株中表达水平极低,因此利用GAL1-10启动子在酵母中过量表达SWI5基因产物,并使用蛋白质提取物证明SWI5蛋白在体外与体内转录激活所需的HO基因启动子片段结合。