Swift S, McGraw P
Department of Biological Sciences, University of Maryland (UMBC), Catonsville 21228, USA.
Nucleic Acids Res. 1995 Apr 25;23(8):1426-33. doi: 10.1093/nar/23.8.1426.
A dominant allele of the INO1 locus, INO1-100, does not require the positive regulators encoded by INO2 and INO4 for expression. Sequence analysis showed that INO1-100 had a 239 bp deletion in the INO1 promoter. INO1-100 suppressed the inositol auxotrophy of ino2, ino4, swi1, swi2 and swi3 mutants. Transcription of INO1-100 was constitutive and independent of these regulators. A 20 bp deletion from -247 to -228 caused a similar phenotype. A 38 bp deletion from -245 to -208 suppressed the inositol auxotrophy of an ino2 mutant, but not an ino4 mutant, indicating that Ino2p and Ino4p may function alone as well as in a complex. A 40 bp deletion from -287 to -248 that removed a URS1 site caused constitutive transcription that required INO2 and INO4. A deletion from -167 to -128 suppressed the inositol auxotrophy of swi, ino2 and ino4 mutants, indicating the presence of a previously unidentified URS1. Mutation of the specific negative regulator of phospholipid synthesis encoded by OPI1 suppressed the inositol auxotrophy of swi2 mutants. This study indicates that negative regulation of INO1 is chromatin mediated and provides in vivo information on the interaction of both general and specific regulatory factors that function to accomplish negative and positive regulation of the INO1 promoter in response to inositol.
INO1基因座的显性等位基因INO1 - 100的表达不需要INO2和INO4编码的正向调节因子。序列分析表明,INO1 - 100在INO1启动子中有一个239 bp的缺失。INO1 - 100抑制了ino2、ino4、swi1、swi2和swi3突变体的肌醇营养缺陷型。INO1 - 100的转录是组成型的,且独立于这些调节因子。从 - 247到 - 228缺失20 bp会导致类似的表型。从 - 245到 - 208缺失38 bp可抑制ino2突变体的肌醇营养缺陷型,但不能抑制ino4突变体,这表明Ino2p和Ino4p可能单独发挥作用,也可能在复合物中发挥作用。从 - 287到 - 248缺失40 bp(去除了一个URS1位点)导致组成型转录,这需要INO2和INO4。从 - 167到 - 128的缺失抑制了swi、ino2和ino4突变体的肌醇营养缺陷型,表明存在一个先前未鉴定的URS1。由OPI1编码的磷脂合成特异性负调节因子的突变抑制了swi2突变体的肌醇营养缺陷型。这项研究表明,INO1的负调控是由染色质介导的,并提供了体内关于一般和特异性调节因子相互作用的信息,这些调节因子在响应肌醇时对INO1启动子进行负向和正向调节。