Phenotypic, proteomic, and genomic characterization of a putative ABC-transporter permease involved in Listeria monocytogenes biofilm formation.
Phenotypic, proteomic, and genomic characterization of a putative ABC-transporter permease involved in Listeria monocytogenes biofilm formation.
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DOI：
10.1089/fpd.2010.0697
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发表时间：
2011-03
影响因子：
2.8
通讯作者：
Xin-na Zhu;Wei-bing Liu;R. Lametsch;F. Aarestrup;Chunlei Shi;Q. She;Xianming Shi;S. Knøchel
中科院分区：
文献类型：
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作者：
Xin-na Zhu;Wei-bing Liu;R. Lametsch;F. Aarestrup;Chunlei Shi;Q. She;Xianming Shi;S. Knøchel
食源性病原体单核增生李斯特菌能够在食品加工环境中形成生物膜。之前，我们曾报道过一个lm。G_1771基因（编码假定的abc转运体渗透酶）参与利用LM-49对单核增生乳杆菌生物膜形成的负调控，LM-49是Tn917诱变分离的生物膜增强突变体（AEM 2008 p.7675-7683）。本研究通过表型、蛋白质组学和基因组学分析对这种abc转运蛋白渗透酶在单核增生乳杆菌生物膜形成中的可能作用进行了表征。G_1771基因缺失（Δ1771）。通过结晶紫染色试验，Δ1771突变体表现出与LM-49菌株相同的增强的生物膜形成能力。DNA微阵列和双向凝胶电泳分别在Δ1771中发现了49个和11个差异表达的基因或蛋白。转录组学研究表明。G_1771可能在调节参与生物膜形成的候选基因中发挥重要作用，如编码细胞表面蛋白（Dlt）、细胞表面锚定蛋白（SrtA）和转录调节因子（GntR）的基因，这些基因对单核增生乳杆菌的生物膜形成起负调控作用。突变体Δ1771对Triton X-100更敏感，而对阳离子抗生素的抗性更弱，这可能是由于该突变体中dlt操纵子的下调，以及dlt与d -丙氨酸残基结合到脂质磷壁酸中，导致磷壁酸带正电荷。因此,lm。G_1771被认为参与了生物膜形成的负调控，本研究结果为lm调控生物膜形成提供了可能的分子机制。单核增生乳杆菌中G_1771。
The foodborne pathogen Listeria monocytogenes is able to form biofilms in food processing environments. Previously, we have reported that an lm.G_1771 gene (encoding a putative ABC-transporter permease) was involved in negative regulation of L. monocytogenes biofilm formation using LM-49, a biofilm-enhanced mutant isolated on Tn917 mutagenesis (AEM 2008 p.7675-7683). Here, the possible action of this ABC-transporter permease in L. monocytogenes biofilm formation was characterized by phenotypic, proteomic, and genomic analyses using an lm.G_1771 gene deletant (Δ1771). The Δ1771 mutant exhibited the same enhanced ability for biofilm formation as the LM-49 strain using a crystal violet staining assay. DNA microarrays and two-dimensional gel electrophoresis revealed 49 and 11 differentially expressed (twofold or more) genes or proteins in Δ1771, respectively. The transcriptomics study indicated that lm.G_1771 could play a vital role in regulating candidate genes involved in biofilm formation such as genes encoding cell surface proteins (Dlt), cell surface anchor proteins (SrtA), and transcriptional regulators (GntR) contributing to negative regulation of biofilm formation by L. monocytogenes. The mutant Δ1771 was more sensitive to Triton X-100 and less resistant to cationic antibiotics, which might be explained by the down-regulation of dlt operon in this deletant and the fact that dlt involves the incorporation of D-alanine residues into lipoteichoic acids, resulting in a positive net charge on the teichoic acids. Therefore, lm.G_1771 is considered to be involved in negative regulation of biofilm formation, and the results from this work provide a possible molecular mechanism of biofilm formation regulated by lm.G_1771 in L. monocytogenes.