Weber Barbara, Croxatto Antony, Chen Chang, Milton Debra L
Department of Molecular Biology, Umeå University, S-901 87 Umeå, Sweden.
Microbiology (Reading). 2008 Mar;154(Pt 3):767-780. doi: 10.1099/mic.0.2007/014167-0.
In vibrios, regulation of the Vibrio harveyi-like LuxR transcriptional activators occurs post-transcriptionally via small regulatory RNAs (sRNAs) that destabilize the luxR mRNA at a low cell population, eliminating expression of LuxR. Expression of the sRNAs is modulated by the vibrio quorum-sensing phosphorelay systems. However, vanT mRNA, which encodes a LuxR homologue in Vibrio anguillarum, is abundant at low and high cell density, indicating that VanT expression may be regulated via additional mechanisms. In this study, Western analyses showed that VanT was expressed throughout growth with a peak of expression during late exponential growth. VanO induced partial destabilization of vanT mRNA via activation of at least one Qrr sRNA. Interestingly, the sigma factor RpoS significantly stabilized vanT mRNA and induced VanT expression during late exponential growth. This induction was in part due to RpoS repressing expression of Hfq, an RNA chaperone. RpoS is not part of the quorum-sensing regulatory cascade since RpoS did not regulate expression or activity of VanO, and RpoS was not regulated by VanO or VanT. VanT and RpoS were needed for survival following UV irradiation and for pigment and metalloprotease production, suggesting that RpoS works with the quorum-sensing systems to modulate expression of VanT, which regulates survival and stress responses.
在弧菌中,类哈维氏弧菌LuxR转录激活因子的调控发生在转录后,通过小调控RNA(sRNA)在低细胞群体中使luxR mRNA不稳定,从而消除LuxR的表达。sRNA的表达受弧菌群体感应磷酸化信号转导系统调节。然而,在鳗弧菌中编码LuxR同源物的vanT mRNA在低细胞密度和高细胞密度时都很丰富,这表明VanT的表达可能通过其他机制调控。在本研究中,蛋白质免疫印迹分析表明,VanT在整个生长过程中都有表达,在指数生长后期表达达到峰值。VanO通过激活至少一种Qrr sRNA诱导vanT mRNA部分不稳定。有趣的是,σ因子RpoS在指数生长后期显著稳定vanT mRNA并诱导VanT表达。这种诱导部分是由于RpoS抑制了RNA伴侣蛋白Hfq的表达。RpoS不是群体感应调控级联的一部分,因为RpoS不调节VanO的表达或活性,且RpoS不受VanO或VanT调节。UV照射后存活以及色素和金属蛋白酶产生都需要VanT和RpoS,这表明RpoS与群体感应系统协同调节VanT的表达,而VanT调节存活和应激反应。