Fyfe J A, Strugnell R A, Davies J K
Department of Microbiology, Monash University, Clayton, Victoria, Australia.
Gene. 1993 Jan 15;123(1):45-50. doi: 10.1016/0378-1119(93)90537-d.
The pilE gene from Neisseria gonorrhoeae, unlike other type-4 pilin-encoding genes, is well expressed in Escherichia coli. Two putative promoters have been implicated in the transcription of this gene. Besides the -24/-12 promoter used to transcribe type-4 pilin-encoding genes in most species, the consensus sequence for a conventional promoter is also present. The two promoters overlap and would have almost identical transcription start points (tsp). Transcription from a -24/-12 promoter should be abolished in an E. coli rpoN mutant. A recombinant plasmid carrying pilE could not be transformed into such a mutant, apparently because the synthesis of the N-terminal hydrophobic domain of pilin is lethal to the rpoN mutant. This suggests that pilE is expressed at a higher level in an rpoN mutant than it is in a wild-type (wt) strain of E. coli. This suggestion was confirmed by constructing fusions between the pilE promoter region and a promoter-less cat gene. We suggest that the conventional promoter is primarily responsible for the transcription of pilE, but that the binding of the RpoN sigma factor partially represses transcription of this gene in wt strains. In an rpoN mutant, the repression is removed and transcription occurs at a level that is lethal to the mutant host.
淋病奈瑟菌的pilE基因与其他4型菌毛编码基因不同,它在大肠杆菌中能良好表达。该基因的转录涉及两个推定的启动子。除了大多数物种中用于转录4型菌毛编码基因的-24/-12启动子外,还存在一个常规启动子的共有序列。这两个启动子相互重叠,转录起始点(tsp)几乎相同。在大肠杆菌rpoN突变体中,来自-24/-12启动子的转录应被消除。携带pilE的重组质粒无法转化到这样的突变体中,显然是因为菌毛N端疏水区的合成对rpoN突变体是致死性的。这表明pilE在rpoN突变体中的表达水平高于在大肠杆菌野生型(wt)菌株中的表达水平。通过构建pilE启动子区域与无启动子cat基因之间的融合体,这一推测得到了证实。我们认为常规启动子主要负责pilE的转录,但RpoN σ因子的结合在wt菌株中会部分抑制该基因的转录。在rpoN突变体中,这种抑制作用被消除,转录以对突变宿主致死的水平发生。