Hoang Viet, Bini Elisabetta, Dixit Vidula, Drozda Melissa, Blum Paul
George Beadle Center for Genetics, University of Nebraska, Lincoln, Nebraska 68588-0666, USA.
Genetics. 2004 Aug;167(4):1563-72. doi: 10.1534/genetics.103.024380.
The archaeon Sulfolobus solfataricus uses a catabolite repression-like system to control production of several glycoside hydrolases. To better understand this regulatory system, studies of the regulation of expression of the beta-glycosidase gene (lacS) were conducted. Expression of lacS varies in response to medium composition and to mutations at an unlinked gene called car. Despite gene overlap, expression of the lacS promoter proximal gene, SSO3017, exhibited coregulation but not cotranscription with lacS. Measurements of mRNA half-life excluded differential stability as a factor in lacS regulation. Chromosomal repositioning by homologous recombination of a lacS deletion series clarified critical cis-acting sequences required for lacS regulation. lacS repositioned at amyA exhibited increased lacS expression and compromised the response to medium composition independently of lacS 5' flanking sequence composition. In contrast, regulation of lacS by the car mutation was dependent on sequences upstream of the archaeal TATA box. Expression of a promoter fusion between lacS and the car-independent malA promoter integrated either at amyA or at the natural lacS locus was insensitive to the allelic state of car. In contrast, the promoter fusion retained a response to medium composition only at the lacS locus. These results indicate that car acts at the lacS promoter and that the response to medium composition involves locus-specific sequences exclusive of those present 5' to lacS or within the lacS transcription unit.
嗜热栖热放线菌利用一种类似分解代谢物阻遏的系统来控制几种糖苷水解酶的产生。为了更好地理解这个调控系统,我们对β-糖苷酶基因(lacS)的表达调控进行了研究。lacS的表达会根据培养基成分以及一个名为car的非连锁基因突变而发生变化。尽管基因存在重叠,但lacS启动子近端基因SSO3017的表达表现出协同调控,但与lacS不共转录。mRNA半衰期的测量排除了差异稳定性作为lacS调控因素的可能性。通过lacS缺失系列的同源重组进行染色体重新定位,明确了lacS调控所需的关键顺式作用序列。定位在amyA处的lacS表现出lacS表达增加,并且独立于lacS 5'侧翼序列组成而损害了对培养基成分的反应。相比之下,car突变对lacS的调控依赖于古菌TATA框上游的序列。整合在amyA或天然lacS位点的lacS与不依赖car的malA启动子之间的启动子融合表达对car的等位基因状态不敏感。相反,启动子融合仅在lacS位点保留了对培养基成分的反应。这些结果表明,car作用于lacS启动子,并且对培养基成分的反应涉及特定于位点的序列,这些序列不包括lacS 5'端或lacS转录单元内的序列。