Rieu Aurélie, Weidmann Stéphanie, Garmyn Dominique, Piveteau Pascal, Guzzo Jean
UMR 1229 Microbiologie du Sol et de l'Environnement, Université de Bourgogne, INRA, F-21000 Dijon, France.
Appl Environ Microbiol. 2007 Oct;73(19):6125-33. doi: 10.1128/AEM.00608-07. Epub 2007 Aug 3.
In this study, we investigated the agrBDCA operon in the pathogenic bacterium Listeria monocytogenes EGD-e. In-frame deletion of agrA and agrD resulted in an altered adherence and biofilm formation on abiotic surfaces, suggesting the involvement of the agr system of L. monocytogenes during the early stages of biofilm formation. Real-time PCR experiments indicated that the transcript levels of agrBDCA depended on the stage of biofilm development, since the levels were lower after the initial attachment period than during biofilm growth, whereas transcription during planktonic growth was not growth phase dependent. The mRNA quantification data also suggested that the agr system was autoregulated and pointed to a differential expression of the agr genes during sessile and planktonic growth. Although the reverse transcription-PCR experiments revealed that the four genes were transcribed as a single messenger, chemical half-life and 5' RACE (rapid amplification of cDNA ends) experiments indicated that the full size transcript underwent cleavage followed by degradation of the agrC and agrA transcripts, which suggests a complex regulation of agr transcription.
在本研究中,我们对致病性细菌单核细胞增生李斯特菌EGD-e中的agrBDCA操纵子进行了研究。agrA和agrD的框内缺失导致在非生物表面上的黏附及生物膜形成发生改变,这表明单核细胞增生李斯特菌的agr系统在生物膜形成的早期阶段发挥作用。实时PCR实验表明,agrBDCA的转录水平取决于生物膜发育阶段,因为在初始附着期后其水平低于生物膜生长期间,而浮游生长期间的转录不依赖于生长阶段。mRNA定量数据还表明,agr系统存在自我调节,并且表明agr基因在固着生长和浮游生长期间存在差异表达。尽管逆转录PCR实验显示这四个基因作为单个信使进行转录,但化学半衰期和5' RACE(cDNA末端快速扩增)实验表明,全长转录本会发生切割,随后agrC和agrA转录本会降解,这表明agr转录存在复杂的调控机制。