Wu S S, Kaiser D
Department of Biochemistry, Stanford University School of Medicine, California 94305, USA.
J Bacteriol. 1997 Dec;179(24):7748-58. doi: 10.1128/jb.179.24.7748-7758.1997.
Type IV pili are required for social gliding motility in Myxococcus xanthus. In this work, the expression of pilin (the pilA gene product) during vegetative growth and fruiting-body development was examined. A polyclonal antibody against the pilA gene product (prepilin) was prepared, along with a pilA-lacZ fusion, and was used to assay expression of pilA in M. xanthus in different mutant backgrounds. pilA expression required the response regulator pilR but was negatively regulated by the putative sensor kinase pilS. pilA expression did not require pilB, pilC, or pilT. pilA was also autoregulated; a mutation which altered an invariant glutamate five residues from the presumed prepilin processing site eliminated this autoregulation, as did a deletion of the pilA gene. Primer extension and S1 nuclease analysis identified a sigma54 promoter upstream of pilA, consistent with the homology of pilR to the NtrC family of response regulators. Expression of pilA was found to be developmentally regulated; however, the timing of this expression pattern was not entirely dependent on pilS or pilR. Finally, pilA expression was induced by high nutrient concentrations, an effect that was also not dependent on pilS or pilR.
IV型菌毛是黄色黏球菌进行群体滑动运动所必需的。在这项研究中,我们检测了菌毛蛋白(pilA基因产物)在营养生长和子实体发育过程中的表达情况。制备了一种针对pilA基因产物(前菌毛蛋白)的多克隆抗体,以及一个pilA - lacZ融合体,并用于检测黄色黏球菌在不同突变背景下pilA的表达。pilA的表达需要应答调节因子pilR,但受到假定的传感激酶pilS的负调控。pilA的表达不需要pilB、pilC或pilT。pilA也存在自我调节;一个改变了假定前菌毛蛋白加工位点上游五个不变谷氨酸残基的突变消除了这种自我调节,pilA基因的缺失也有同样的效果。引物延伸和S1核酸酶分析确定了pilA上游的一个sigma54启动子,这与pilR与NtrC家族应答调节因子的同源性一致。发现pilA的表达受到发育调控;然而,这种表达模式的时间并不完全依赖于pilS或pilR。最后,高营养浓度可诱导pilA的表达,这种效应也不依赖于pilS或pilR。