Bassler B L, Wright M, Silverman M R
Agouron Institute, La Jolla, California 92037.
Mol Microbiol. 1994 May;12(3):403-12. doi: 10.1111/j.1365-2958.1994.tb01029.x.
Density-dependent expression of luminescence in Vibrio harveyi is regulated by the concentration of extracellular signal molecules (autoinducers) in the culture medium. A recombinant clone that restored function to one class of spontaneous dim mutants was found to encode a function required for the density-dependent response. Transposon Tn5 insertions in the recombinant clone were isolated, and the mutations were transferred to the genome of V. harveyi for examination of mutant phenotypes. Expression of luminescence in V. harveyi strains with transposon insertions in one locus, luxO, was independent of the density of the culture and was similar in intensity to the maximal level observed in wild-type bacteria. Sequence analysis of luxO revealed one open reading frame that encoded a protein, LuxO, similar in amino acid sequence to the response regulator domain of the family of two-component, signal transduction proteins. The constitutive phenotype of LuxO- mutants indicates that LuxO acts negatively to control expression of luminescence, and relief of repression by LuxO in the wild type could result from interactions with other components in the Lux signalling system.
哈氏弧菌中发光的密度依赖性表达受培养基中细胞外信号分子(自诱导物)浓度的调节。发现一个能恢复一类自发暗突变体功能的重组克隆编码了密度依赖性反应所需的一种功能。分离出重组克隆中的转座子Tn5插入片段,并将突变转移到哈氏弧菌基因组中以检测突变表型。在一个位点luxO中有转座子插入的哈氏弧菌菌株中,发光表达与培养密度无关,其强度与野生型细菌中观察到的最大水平相似。对luxO的序列分析揭示了一个开放阅读框,其编码一种蛋白质LuxO,该蛋白质的氨基酸序列与双组分信号转导蛋白家族的应答调节结构域相似。LuxO突变体的组成型表型表明LuxO对发光表达起负调控作用,野生型中LuxO抑制作用的解除可能是由于与Lux信号系统中的其他组分相互作用所致。