Shadel G S, Baldwin T O
Department of Biochemistry and Biophysics, Texas A&M University, College Station.
J Bacteriol. 1991 Jan;173(2):568-74. doi: 10.1128/jb.173.2.568-574.1991.
Regulation of the genes required for bioluminescence in the marine bacterium Vibrio fischeri (the lux regulon) is a complex process requiring coordination of several systems. The primary level of regulation is mediated by a positive regulatory protein, LuxR, and a small diffusible molecule, N-(3-oxo-hexanoyl)-homoserine lactone, termed autoinducer. Transcription of the luxR gene, which encodes the regulatory protein, is positively regulated by the cyclic AMP-CAP system. The lux regulon of V. fischeri consists of two divergently transcribed operons designated operonL and operonR. Transcription of the rightward operon (operonR; luxICDABE), consisting of the genes required for autoinducer synthesis (luxI) and light production (luxCDABE), is activated by LuxR in an autoinducer-dependent fashion. The leftward operon (operonL) consists of a single known gene, luxR. The LuxR protein has also been shown to decrease transcription of operonL through an autoinducer-dependent mechanism, thereby negatively regulating its own synthesis. In this paper we demonstrate that the autoinducer-dependent repression of operonL transcription requires not only LuxR but also DNA sequences within operonR which occur upstream of the promoter for operonL. In the absence of these DNA sequences, the LuxR protein causes an autoinducer-dependent activation of transcription of operonL. The lux operator, located in the control region between the two operons, was required for both the positive and negative autoinducer-dependent responses. By titration of high levels of LuxR supplied in trans with synthetic autoinducer, we found that low levels of autoinducer could elicit a positive response even in the presence of the negative-acting DNA sequences, while higher levels of autoinducer resulted in a negative response. Without these DNA sequences in operonR, LuxR and autoinducer stimulated transcription regardless of the level of autoinducer. These results suggest that a switch between stimulation and repression of operonL transcription is mediated by the levels of the LuxR-autoinducer complex, which in these experiments reflects the level of autoinducer in the growth medium.
费氏弧菌(lux操纵子)中生物发光所需基因的调控是一个复杂的过程,需要多个系统的协调。主要的调控水平由一种正向调控蛋白LuxR和一种小的可扩散分子N-(3-氧代己酰基)-高丝氨酸内酯介导,后者被称为自诱导物。编码调控蛋白的luxR基因的转录受环腺苷酸-分解代谢物激活蛋白(cAMP-CAP)系统的正向调控。费氏弧菌的lux操纵子由两个反向转录的操纵子组成,分别称为操纵子L和操纵子R。由自诱导物合成所需基因(luxI)和发光所需基因(luxCDABE)组成的向右操纵子(操纵子R;luxICDABE)的转录,以自诱导物依赖的方式被LuxR激活。向左操纵子(操纵子L)由一个已知基因luxR组成。LuxR蛋白也已被证明通过一种自诱导物依赖的机制降低操纵子L的转录,从而对其自身的合成进行负调控。在本文中,我们证明操纵子L转录的自诱导物依赖抑制不仅需要LuxR,还需要操纵子R中位于操纵子L启动子上游的DNA序列。在没有这些DNA序列的情况下,LuxR蛋白会导致操纵子L转录的自诱导物依赖激活。位于两个操纵子之间控制区域的lux操纵基因,对于正向和负向的自诱导物依赖反应都是必需的。通过用合成自诱导物滴定反式提供的高水平LuxR,我们发现即使存在负向作用的DNA序列,低水平的自诱导物也能引发正向反应,而较高水平的自诱导物则导致负向反应。如果操纵子R中没有这些DNA序列,LuxR和自诱导物会刺激转录,而与自诱导物的水平无关。这些结果表明,操纵子L转录的刺激和抑制之间的转换是由LuxR-自诱导物复合物的水平介导的,在这些实验中,该复合物的水平反映了生长培养基中自诱导物的水平。