Geever R F, Murayama T, Case M E, Giles N H
Proc Natl Acad Sci U S A. 1986 Jun;83(11):3944-8. doi: 10.1073/pnas.83.11.3944.
Transcriptional activation of the Neurospora crassa qa genes normally requires the positive regulatory gene, qa-1F+, whose function is controlled by the inducer quinic acid and by the product of the negative regulatory gene, qa-1S+. The properties of qa-1F+ activator have been examined in transcriptional mutations of the qa-2 structural gene, in which activator-independent transcription of qa-2 (qa-2ai mutants) occurs in strains having a qa-1F- gene. Seven qa-2ai mutants with DNA rearrangements in different 5' regions of qa-2 were analyzed in qa-1F+ strains. In five with rearrangements at position -190 or further upstream, expression of the qa-2 gene was inducible, and induction was accompanied by a change in the initiation site for transcription from position -45, characteristic of constitutive initiation in qa-2ai mutants to position +1, characteristic of the induced wild type. In two mutants with breakpoints at positions -86 and -53, qa-2 transcription initiated from upstream sequences within the rearrangements but not at the +1 site, and qa-2 expression was noninducible. The results indicate that (i) sequences between positions -190 and -86 are required for positive control of initiation at position +1, and (ii) negative control does not require sequences downstream of position -86. Additional evidence suggests that the product of the qa-1F+ gene in the noninduced state may also interact with distal upstream sequences positioned midway between divergently transcribed qa genes.
粗糙脉孢菌qa基因的转录激活通常需要正向调节基因qa - 1F +,其功能受诱导物奎尼酸和负向调节基因qa - 1S +产物的控制。已在qa - 2结构基因的转录突变体中研究了qa - 1F +激活剂的特性,在具有qa - 1F -基因的菌株中发生了qa - 2的独立于激活剂的转录(qa - 2ai突变体)。在qa - 1F +菌株中分析了七个在qa - 2不同5'区域发生DNA重排的qa - 2ai突变体。在五个在-190位置或更上游发生重排的突变体中,qa - 2基因的表达是可诱导的,并且诱导伴随着转录起始位点从-45位置的变化,-45位置是qa - 2ai突变体组成型起始的特征,变为+1位置,+1位置是诱导型野生型的特征。在两个断点位于-86和-53位置的突变体中,qa - 2转录从重排内的上游序列起始,但不在+1位点起始,并且qa - 2表达是不可诱导的。结果表明:(i)-190和-86位置之间的序列是在+1位置起始进行正向控制所必需的;(ii)负向控制不需要-86位置下游的序列。其他证据表明,处于未诱导状态的qa - 1F +基因产物也可能与位于反向转录的qa基因之间中途位置的远端上游序列相互作用。