Clemmer Katy M, Rather Philip N
Laboratories of Microbial Pathogenesis, Veterans Affairs Medical Center, Decatur, GA, USA.
Res Microbiol. 2007 Apr;158(3):295-302. doi: 10.1016/j.resmic.2006.11.010. Epub 2007 Jan 13.
Transposon insertions located 325 and 740 base pairs upstream of the transcriptional start site of the flhDC operon resulted in cells that initiated swarming 1.5h earlier than wild-type and exhibited a 2-2.5-fold greater swarming velocity. These mutants also failed to consolidate (de-differentiate) normally and did not form the characteristic bulls-eye pattern of concentric swarming rings on solid media. The analysis of one mutant (SS-P) with an insertion at -325 revealed that the levels of flhDC mRNA were dramatically higher than wild-type during swarmer cell differentiation and failed to decrease during the consolidation period. However, the start point of flhDC transcription was identical in the SS-P mutant and wild-type cells. The presence of the flhDC upstream region on a high copy plasmid increased swarming motility and expression of a chromosomal flhDC-lacZ fusion, presumably by titrating out a repressor. To identify potential repressors and further define flhDC regulation in P. mirabilis, targeted disruptions were created in the rcsB, ompR, lrhA and hdfR genes, previously demonstrated to repress flhDC in E. coli. Of these mutations, only the loss of rcsB increased swarming and flhDC mRNA accumulation.
位于flhDC操纵子转录起始位点上游325和740个碱基对处的转座子插入导致细胞比野生型提前1.5小时开始群体游动,并且群体游动速度提高了2至2.5倍。这些突变体在固体培养基上也不能正常巩固(去分化),并且没有形成特征性的同心群体游动环的靶心图案。对一个在-325处有插入的突变体(SS-P)的分析表明,在群体游动细胞分化过程中,flhDC mRNA的水平显著高于野生型,并且在巩固期没有下降。然而,SS-P突变体和野生型细胞中flhDC转录的起始点是相同的。高拷贝质粒上flhDC上游区域的存在增加了群体游动能力和染色体flhDC-lacZ融合体的表达,推测是通过滴定出一种阻遏物来实现的。为了鉴定潜在的阻遏物并进一步确定奇异变形杆菌中flhDC的调控,在rcsB、ompR、lrhA和hdfR基因中产生了靶向破坏,这些基因先前已被证明在大肠杆菌中可抑制flhDC。在这些突变中,只有rcsB的缺失增加了群体游动和flhDC mRNA的积累。