Braun Derek C, Stein Daniel C
Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland 20742, USA.
J Bacteriol. 2004 Feb;186(4):1038-49. doi: 10.1128/JB.186.4.1038-1049.2004.
Biosynthesis of the variable core domain of lipooligosaccharide (LOS) in Neisseria gonorrhoeae is mediated by glycosyl transferases encoded by lgtABCDE. Changes within homopolymeric runs within lgtA, lgtC, and lgtD affect the expression state of these genes, with the nature of the LOS expressed determined by the functionality of these genes. However, the mechanism for modulating the amount of multiple LOS chemotypes expressed in a single cell is not understood. Using mutants containing polar disruptions within the lgtABCDE locus, we determined that the expression of this locus is mediated by multiple promoters and that disruption of transcription from these promoters alters the relative levels of simultaneously expressed LOS chemotypes. Expression of the lgtABCDE locus was quantified by using xylE transcriptional fusions, and the data indicate that this locus is transcribed in trace amounts and that subtle changes in transcription result in phenotypic changes. By using rapid amplification of 5' cDNA ends, transcriptional start sites and promoter sequences were identified within lgtABCDE. Most of these promoters possessed 50 to 67% homology with the consensus gearbox promoter sequence of Escherichia coli.
淋病奈瑟菌脂寡糖(LOS)可变核心结构域的生物合成由lgtABCDE编码的糖基转移酶介导。lgtA、lgtC和lgtD中同聚物序列内的变化会影响这些基因的表达状态,所表达的LOS的性质由这些基因的功能决定。然而,调节单个细胞中多种LOS化学型表达量的机制尚不清楚。利用在lgtABCDE基因座内含有极性破坏的突变体,我们确定该基因座的表达由多个启动子介导,并且这些启动子转录的破坏会改变同时表达的LOS化学型的相对水平。通过使用xylE转录融合对lgtABCDE基因座的表达进行定量,数据表明该基因座以微量转录,转录中的细微变化会导致表型变化。通过使用5' cDNA末端的快速扩增,在lgtABCDE内鉴定了转录起始位点和启动子序列。这些启动子中的大多数与大肠杆菌的共有齿轮箱启动子序列具有50%至67%的同源性。