Geever R F, Case M E, Tyler B M, Buxton F, Giles N H
Proc Natl Acad Sci U S A. 1983 Dec;80(23):7298-302. doi: 10.1073/pnas.80.23.7298.
Expression of the qa-2 gene of Neurospora crassa normally requires a functional activator protein encoded by qa-1F. Twelve transcriptional mutants of the qa-2 gene have been isolated in qa-1F- strains, and these allow partial expression of qa-2 (1-45% of induced wild type) in the absence of functional activator protein. All 12 mutants have been characterized by genomic (Southern) blot hybridization and the DNAs of 5 have been cloned and sequenced. Eight mutations consist of large DNA rearrangements within a 500-base-pair region 5' to the qa-2 gene. One large rearrangement mutation, located 378 base pairs before the normal site of transcription initiation, causes exceptional levels of qa-2 transcription (45% of induced wild type) from near the normal initiation site. Two of the other four mutations cloned involve tandem duplications (68 and 84 base pairs) of the same upstream region (centered at nucleotide - 145), and two involve "point" mutations (at nucleotides -200 and -95) that closely flank the duplicated region. With one possible exception, none of the mutations appears to involve changes directly associated with RNA polymerase II binding and hence they differ from analogous mutations in comparable prokaryotic systems. The overall results suggest that at least some of the large DNA rearrangement mutations may be acting as upstream activator elements, possibly by juxtaposing enhancer-like sequences, whereas the duplications and point mutations may define a region of qa-2 regulation, for instance at the level of RNA polymerase II access.
粗糙脉孢菌的qa - 2基因的表达通常需要由qa - 1F编码的功能性激活蛋白。在qa - 1F -菌株中分离出了12个qa - 2基因的转录突变体,这些突变体在没有功能性激活蛋白的情况下能使qa - 2部分表达(为诱导型野生型的1 - 45%)。所有12个突变体都通过基因组(Southern)印迹杂交进行了表征,其中5个的DNA已被克隆和测序。8个突变由qa - 2基因5'端500个碱基对区域内的大片段DNA重排组成。一个大的重排突变位于转录起始正常位点之前378个碱基对处,导致从正常起始位点附近出现异常高水平的qa - 2转录(为诱导型野生型的45%)。克隆的其他4个突变中有两个涉及同一上游区域(以核苷酸 - 145为中心)的串联重复(68和84个碱基对),另外两个涉及紧密位于重复区域两侧的“点”突变(在核苷酸 - 200和 - 95处)。除了一个可能的例外,这些突变似乎都不涉及与RNA聚合酶II结合直接相关的变化,因此它们与可比原核系统中的类似突变不同。总体结果表明,至少一些大片段DNA重排突变可能作为上游激活元件起作用,可能是通过并列类似增强子的序列,而重复和点突变可能定义了qa - 2的一个调控区域,例如在RNA聚合酶II可及性水平上。