Tomich Mladen, Mohr Christian D
Department of Microbiology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
J Bacteriol. 2004 Feb;186(4):1009-20. doi: 10.1128/JB.186.4.1009-1020.2004.
Burkholderia cenocepacia is an important member of the Burkholderia cepacia complex, a group of closely related bacteria that inhabits a wide variety of environmental niches in nature and that also colonizes the lungs of compromised humans. Certain strains of B. cenocepacia express peritrichous adherence organelles known as cable pili, thought to be important in the colonization of the lower respiratory tract. The genetic locus required for cable pilus biogenesis is comprised of at least five genes, designated cblB, cblA, cblC, cblD, and cblS. In this study a transcriptional analysis of cbl gene expression was undertaken. The principal promoter, located upstream of the cbl locus, was identified and characterized. By using lacZ transcriptional fusions, the effects of multiple environmental cues on cbl gene expression were examined. High osmolarity, temperature of 37 degrees C, acidic pH, and low iron bioavailability were found to induce cbl gene expression. Northern hybridization analysis of the cbl locus identified a single, stable transcript corresponding to cblA, encoding the major pilin subunit. Transcriptional fusion studies combined with reverse transcription-PCR analysis indicated that the stable cblA transcript is the product of an mRNA processing event. This event may ensure high levels of expression of the major pilin, relative to other components of the assembly pathway. Our findings lend further insight into the control of cable pilus biogenesis in B. cenocepacia and provide evidence for regulation of cbl gene expression on both the transcriptional and posttranscriptional levels.
洋葱伯克霍尔德菌是伯克霍尔德菌复合体的重要成员,该复合体是一组密切相关的细菌,它们存在于自然界的各种环境生态位中,也可定殖于免疫力低下人群的肺部。某些洋葱伯克霍尔德菌菌株表达周生附着细胞器,即所谓的束状菌毛,被认为在呼吸道下段定殖中起重要作用。束状菌毛生物合成所需的基因座至少由五个基因组成,分别命名为cblB、cblA、cblC、cblD和cblS。在本研究中,对cbl基因表达进行了转录分析。鉴定并表征了位于cbl基因座上游的主要启动子。通过使用lacZ转录融合,研究了多种环境信号对cbl基因表达的影响。发现高渗透压、37摄氏度的温度、酸性pH值和低铁生物利用率可诱导cbl基因表达。对cbl基因座的Northern杂交分析鉴定出一个对应于cblA的单一稳定转录本,cblA编码主要菌毛蛋白亚基。转录融合研究与逆转录PCR分析相结合表明,稳定的cblA转录本是mRNA加工事件的产物。该事件可能确保主要菌毛蛋白相对于组装途径的其他组分具有高水平的表达。我们的研究结果进一步深入了解了洋葱伯克霍尔德菌中束状菌毛生物合成的调控,并为cbl基因在转录和转录后水平上的表达调控提供了证据。