Blokesch Melanie, Schoolnik Gary K
Department of Microbiology and Immunology, Beckman Center B237, 279 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305, USA.
J Bacteriol. 2008 Nov;190(21):7232-40. doi: 10.1128/JB.00959-08. Epub 2008 Aug 29.
Free extracellular DNA is abundant in many aquatic environments. While much of this DNA will be degraded by nucleases secreted by the surrounding microbial community, some is available as transforming material that can be taken up by naturally competent bacteria. One such species is Vibrio cholerae, an autochthonous member of estuarine, riverine, and marine habitats and the causative agent of cholera, whose competence program is induced after colonization of chitin surfaces. In this study, we investigate how Vibrio cholerae's two extracellular nucleases, Xds and Dns, influence its natural transformability. We show that in the absence of Dns, transformation frequencies are significantly higher than in its presence. During growth on a chitin surface, an increase in transformation efficiency was found to correspond in time with increasing cell density and the repression of dns expression by the quorum-sensing regulator HapR. In contrast, at low cell density, the absence of HapR relieves dns repression, leading to the degradation of free DNA and to the abrogation of the transformation phenotype. Thus, as cell density increases, Vibrio cholerae undergoes a switch from nuclease-mediated degradation of extracellular DNA to the uptake of DNA by bacteria induced to a state of competence by chitin. Taken together, these results suggest the following model: nuclease production by low-density populations of V. cholerae might foster rapid growth by providing a source of nucleotides for the repletion of nucleotide pools. In contrast, the termination of nuclease production by static, high-density populations allows the uptake of intact DNA and coincides with a phase of potential genome diversification.
游离的细胞外DNA在许多水生环境中都很丰富。虽然这些DNA中的大部分会被周围微生物群落分泌的核酸酶降解,但仍有一些可作为转化物质,能被具有天然感受态的细菌摄取。霍乱弧菌就是这样一种细菌,它是河口、河流和海洋栖息地的本地成员,也是霍乱的病原体,其感受态程序在定殖于几丁质表面后被诱导。在本研究中,我们探究了霍乱弧菌的两种细胞外核酸酶Xds和Dns如何影响其天然转化能力。我们发现,在没有Dns的情况下,转化频率显著高于有Dns时。在几丁质表面生长期间,转化效率的提高被发现与细胞密度的增加以及群体感应调节因子HapR对dns表达的抑制在时间上相对应。相反,在低细胞密度下,HapR的缺失会解除对dns的抑制,导致游离DNA的降解并消除转化表型。因此,随着细胞密度的增加,霍乱弧菌经历了从核酸酶介导的细胞外DNA降解到由几丁质诱导进入感受态的细菌摄取DNA的转变。综上所述,这些结果提示了以下模型:低密度霍乱弧菌群体产生核酸酶可能通过提供核苷酸来源以补充核苷酸库来促进快速生长。相反,静止的高密度群体停止产生核酸酶则允许摄取完整DNA,这与潜在的基因组多样化阶段相吻合。