Spencer Matthew R, Beare Paul A, Lamont Iain L
Department of Biochemistry, University of Otago, Dunedin 9054, New Zealand.
J Bacteriol. 2008 Jul;190(14):4865-9. doi: 10.1128/JB.01998-07. Epub 2008 May 23.
Alternative sigma factor proteins enable transcription of specific sets of genes in bacterial cells. Their activities can be controlled by posttranslational mechanisms including inhibition by antisigma proteins and proteolytic degradation. PvdS is an alternative sigma factor that is required for expression of genes involved in synthesis of a siderophore, pyoverdine, by Pseudomonas aeruginosa. In the absence of pyoverdine, the activity of PvdS is inhibited by a membrane-spanning antisigma factor, FpvR. Inhibition is relieved by a cell surface signaling pathway. In this pathway, a combination of pyoverdine and a cell surface receptor protein, FpvA, suppresses the antisigma activity of FpvR, enabling transcription of PvdS-dependent genes. In this research, we investigated proteolytic degradation of PvdS in response to the signaling pathway. Proteolysis of PvdS was observed in strains of P. aeruginosa in which FpvR had anti-sigma factor activity due to the absence of pyoverdine or the FpvA receptor protein or overproduction of FpvR. Suppression of antisigma activity by addition of pyoverdine or through the absence of FpvR prevented detectable proteolysis of PvdS. The amounts of PvdS were less in bacteria in which proteolysis was observed, and reporter gene assays showed that this reduction was not due to decreased expression of PvdS. In wild-type bacteria, there was an average of 730 molecules of PvdS per cell in late exponential growth phase. Our results show that proteolysis and amounts of PvdS are affected by the antisigma factor FpvR and that this activity of FpvR is controlled by the cell surface signaling pathway.
替代σ因子蛋白能够使细菌细胞中特定基因集进行转录。它们的活性可通过翻译后机制来控制,包括受抗σ蛋白的抑制和蛋白水解降解。PvdS是一种替代σ因子,铜绿假单胞菌合成铁载体绿脓菌素所涉及的基因表达需要它。在没有绿脓菌素的情况下,PvdS的活性受到跨膜抗σ因子FpvR的抑制。细胞表面信号通路可解除这种抑制。在该信号通路中,绿脓菌素和细胞表面受体蛋白FpvA的组合抑制了FpvR的抗σ活性,从而使依赖PvdS的基因得以转录。在本研究中,我们调查了PvdS响应信号通路的蛋白水解降解情况。在因缺乏绿脓菌素或FpvA受体蛋白或FpvR过量产生而使FpvR具有抗σ因子活性的铜绿假单胞菌菌株中,观察到了PvdS的蛋白水解现象。添加绿脓菌素或通过缺失FpvR来抑制抗σ活性可防止检测到PvdS的蛋白水解。在观察到蛋白水解的细菌中,PvdS的量较少,报告基因检测表明这种减少并非由于PvdS表达降低所致。在野生型细菌中,指数生长后期每个细胞平均有730个PvdS分子。我们的结果表明,PvdS的蛋白水解和量受抗σ因子FpvR的影响,并且FpvR的这种活性受细胞表面信号通路的控制。