Desai Kunal, Mashburn-Warren Lauren, Federle Michael J, Morrison Donald A
Laboratory for Molecular Biology, Department of Biological Sciences, College of Liberal Arts and Sciences, University of Illinois at Chicago, Chicago, Illinois, USA.
J Bacteriol. 2012 Aug;194(15):3774-80. doi: 10.1128/JB.00337-12. Epub 2012 May 18.
Streptococcus mutans develops competence for genetic transformation in response to regulatory circuits that sense at least two peptide pheromones. One peptide, known as CSP, is sensed by a two-component signal transduction system through a membrane receptor, ComD. The other, derived from the primary translation product ComS, is thought to be sensed by an intracellular receptor, ComR, after uptake by oligopeptide permease. To allow study of this process in a medium that does not itself contain peptides, development of competence was examined in the chemically defined medium (CDM) described by van de Rijn and Kessler (Infect. Immun. 27:444, 1980). We confirmed a previous report that in this medium comS mutants of strain UA159 respond to a synthetic peptide comprising the seven C-terminal residues of ComS (ComS(11-17)) by increasing expression of the alternative sigma factor SigX, which in turn allows expression of competence effector genes. This response provided the basis for a bioassay for the ComS pheromone in the 100 to 1,000 nM range. It was further observed that comS(+) (but not comS mutant) cultures developed a high level of competence in the late log and transition phases of growth in this CDM without the introduction of any synthetic stimulatory peptide. This endogenous competence development was accompanied by extracellular release of one or more signals that complemented a comS mutation at levels equivalent to 1 μM synthetic ComS(11-17).
变形链球菌会响应至少感知两种肽类信息素的调控回路而产生遗传转化能力。一种名为CSP的肽,通过膜受体ComD被双组分信号转导系统感知。另一种源自初级翻译产物ComS,在被寡肽通透酶摄取后,被认为是由细胞内受体ComR感知。为了在本身不含肽的培养基中研究这一过程,我们在van de Rijn和Kessler(《感染与免疫》27:444,1980)描述的化学限定培养基(CDM)中检测了感受态的发育情况。我们证实了之前的一份报告,即在这种培养基中,UA159菌株的comS突变体通过增加替代西格玛因子SigX的表达来响应包含ComS七个C端残基的合成肽(ComS(11 - 17)),这反过来又使得感受态效应基因得以表达。这种反应为100至1000 nM范围内ComS信息素的生物测定提供了基础。进一步观察到,在这种CDM中,comS(+)(而非comS突变体)培养物在对数生长后期和生长转变期能产生高水平的感受态,而无需引入任何合成刺激肽。这种内源性感受态发育伴随着一种或多种信号的细胞外释放,这些信号能在相当于1 μM合成ComS(11 - 17)的水平上互补comS突变。