Baker S M, Masi A, Liu D F, Novitsky B K, Deich R A
Lederle-Praxis Biologics, West Henrietta, New York 14586, USA.
Infect Immun. 1995 Oct;63(10):3920-6. doi: 10.1128/iai.63.10.3920-3926.1995.
The gene products from an 8-kb region adjacent to the 3' end of the ptx operon are required by Bordetella pertussis for the export of pertussis holotoxin. At least one of these gene products (PtlC) is specifically required for the export of assembled holotoxin from the periplasmic space. ptlC mutants exhibit a 20-fold reduction in the amount of holotoxin present in the culture supernatant but have no effect upon the assembly or steady-state level of holotoxin present in the periplasmic space. Impaired export of holotoxin from the ptlC strain blocks expression of toxin at a posttranscriptional level, and wild-type levels of ptx mRNA are detected in the mutant strain. The transcription of ptl is subject to modulation by MgSO4 in the same manner as ptx is; however, in B. pertussis strains containing an E. coli tac promoter in place of the native ptx promoter, wild-type levels of ptx mRNA are present and holotoxin is synthesized and exported even in the presence of MgSO4. Promoter mapping of the region extending from the ptxS3 coding region to the ptlC coding region failed to detect the ptl transcription initiation site. Additional RNase protection experiments with ptx promoter deletion and substitution strains indicate that the ptl operon is transcribed from the ptx promoter as part of a > 11-kb mRNA.
百日咳博德特氏菌输出百日咳全毒素需要ptx操纵子3'端相邻8 kb区域的基因产物。这些基因产物中至少有一种(PtlC)对于将组装好的全毒素从周质空间输出是特异性必需的。ptlC突变体在培养上清液中存在的全毒素量减少了20倍,但对周质空间中存在的全毒素的组装或稳态水平没有影响。来自ptlC菌株的全毒素输出受损在转录后水平阻断了毒素的表达,并且在突变菌株中检测到野生型水平的ptx mRNA。ptl的转录与ptx一样受到MgSO4的调节;然而,在含有大肠杆菌tac启动子代替天然ptx启动子的百日咳博德特氏菌菌株中,即使在存在MgSO4的情况下也存在野生型水平的ptx mRNA,并且全毒素被合成和输出。从ptxS3编码区延伸到ptlC编码区的区域的启动子定位未能检测到ptl转录起始位点。使用ptx启动子缺失和替换菌株进行的额外RNase保护实验表明,ptl操纵子作为> 11 kb mRNA的一部分从ptx启动子转录。