An agr-like two-component regulatory system in Lactobacillus plantarum is involved in production of a novel cyclic peptide and regulation of adherence
An agr-like two-component regulatory system in Lactobacillus plantarum is involved in production of a novel cyclic peptide and regulation of adherence
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DOI：
10.1128/jb.187.15.5224-5235.2005
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发表时间：
2005-08-01
影响因子：
3.2
通讯作者：
de Vos, WM
中科院分区：
文献类型：
--
作者：
Sturme, MHJ;Nakayama, J;de Vos, WM
我们分析了注释的植物乳杆菌WCFS 1基因组上的一个位点，该位点与葡萄球菌agr群体感应系统具有同源性，并将其指定为乳杆菌agr样模块的lam。显示lamBDCA转录物的产生是生长期依赖性的。在粘附试验中对应答调节因子缺陷型突变体（Δ lamA）的分析表明，lam调节L.玻璃表面的植物。使用基于克隆的微阵列对生长的早期、中期和晚期对数期的野生型和DeltalamA菌株进行全局转录分析。值得注意的是，只有一小部分基因在野生型和lamA突变株之间的转录谱中显示出显著差异。微阵列分析证实，lamBDCA是自动调节的，并表明lamA参与调节编码表面多糖，细胞膜蛋白和糖利用蛋白的基因的表达。使用乳链菌肽控制的表达系统过表达编码推定的自身诱导肽前体（LamD）及其加工蛋白（LamB）的lamBD基因，并通过液相色谱/质谱（LC/MS）分析培养上清液以鉴定过量产生的Lam衍生肽。以这种方式，一个环硫内酯五肽，具有类似的葡萄球菌agr系统的自诱导肽的环结构被确定。该肽被命名为LamD 558，其序列（CVGIW）与注释的前体肽序列匹配。通过LC/MS对野生型培养物上清液的时程分析表明，LamD 558产量从对数生长中期到对数生长后期显著增加。这是第一个在非致病性细菌中编码环状硫内酯自诱导肽并参与粘附调节的agr样系统的例子。
We have analyzed a locus on the annotated Lactobacillus plantarum WCFS1 genome that showed homology to the staphylococcal agr quorum-sensing system and designated it lam for Lactobacillus agr-like module. Production of the lamBDCA transcript was shown to be growth phase dependent. Analysis of a response regulator-defective mutant (Delta lamA) in an adherence assay showed that lam regulates adherence of L. plantarum to a glass surface. Global transcription analysis of the wild-type and Delta lamA strains in early, mid-, and late log phase of growth was performed using a clone-based microarray. Remarkably, only a small set of genes showed significant differences in transcription profiles between the wild-type and lamA mutant strains. The microarray analysis confirmed that lamBDCA is autoregulatory and showed that lamA is involved in regulation of expression of genes encoding surface polysaccharides, cell membrane proteins, and sugar utilization proteins. The lamBD genes encoding the putative autoinducing peptide precursor (LamD) and its processing protein (LamB) were overexpressed using the nisin-controlled expression system, and culture supernatants were analyzed by liquid chromatography/mass spectrometry (LC/MS) to identify overproduced Lam-derived peptides. In this way, a cyclic thiolactone pentapeptide that possesses a ring structure similar to those of autoinducing peptides of the staphylococcal agr system was identified. The peptide was designated LamD558, and its sequence (CVGIW) matched the annotated precursor peptide sequence. Time course analysis of wild-type culture supernatants by LC/MS indicated that LamD558 production was increased markedly from mid-log to late log growth phase. This is the first example of an agr-like system in nonpathogenic bacteria that encodes a cyclic thiolactone autoinducing peptide and is involved in regulation of adherence.