Development of novel anti-microbial drugs targeting quorum sensing in Gram-positive bacteria

开发针对革兰氏阳性菌群体感应的新型抗菌药物

• 批准号: 17580068
• 负责人: NAKAYAMA Jiro
• 金额: $ 2.37万
• 依托单位: KYUSHU UNIVERCITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17580068/
• 关键词: Enterococcus faecalis; Quorum sensing; Screening; Inhibitor; Gelatinase; Virulence factor; Anti-pathogenic compound; Post-antibiotics

The expression of two Enterococcus feacalis virulence-related proteases, gelatinase and serine protease, is positively regulated by a quorum sensing system encoded by the fsr gene cluster. In this system, E. feacalis secretes an autoinducing peptide, gelatinase biosynthesis-activating pheromone(GBAP), which triggers the FsrC-FsrA two component regulatory system controlling the expression of two transcripts, fsrBDC and gelE-sprE. To aim anti-pathogenic compounds targeting cyclic peptide-mediated quorum sensing in Gram-positive bacteria, we have studied on (1) biosynthetic molecular mechanism of cyclic peptide quormone 'GBAP' and (2) screening for inhibitors of GBAP-mediated quorum sensing 'fir regulatory system'.(1) In this study, we demonstrated the existence offsrD, encoding the GBAP propeptide, which is in frame with fsrB but is translated independently of fsrB. It was also demonstrated that FsrB', an FsrD-truncated FsrB, functions as a cysteine protease-like processing enzyme to generate GBAP from FsrD. This revised model is consistent with the staphylococcal agr system.(2) We have screened for inhibitors of the fsr quorum sensing from actinomycetes metabolites, and identified a known peptide antibiotic, siamycin I. Quantitative analysis offsrBDC and gelE-sprE transcript suggested that siamycin I inhibited the GBAP-signaling via FsrC-FsrA two component regulatory system in a noncompetitive maner. The sublethal concentration of siamysin I also attenuated biofilm formation. Overrall, siamycin I would offer a novel means of treating enterococcal infections.

两种粪肠球菌毒力相关蛋白酶（明胶酶和丝氨酸蛋白酶）的表达受到 fsr 基因簇编码的群体感应系统的正向调节。在这个系统中，粪肠球菌分泌一种自诱导肽——明胶酶生物合成激活信息素（GBAP），它触发FsrC-FsrA两部分调节系统，控制两种转录物fsrBDC和gelE-sprE的表达。为了针对革兰氏阳性菌中环肽介导的群体感应的抗病原化合物，我们研究了（1）环肽群体感应“GBAP”的生物合成分子机制和（2）筛选GBAP介导的群体感应“的抑制剂” (1) 在这项研究中，我们证明了offsrD的存在，编码GBAP前肽，它与fsrB在框架内，但翻译独立于fsrB fsrB。还证明 FsrB'（一种 FsrD 截短的 FsrB）可充当半胱氨酸蛋白酶样加工酶，从 FsrD 生成 GBAP。这个修改后的模型与葡萄球菌agr系统一致。（2）我们从放线菌代谢物中筛选了fsr群体感应的抑制剂，并鉴定了一种已知的肽抗生素，siamycin I。srBDC和gelE-sprE转录本的定量分析表明siamycin I以非竞争性方式通过 FsrC-FsrA 两部分调节系统抑制 GBAP 信号传导。亚致死浓度的暹罗霉素 I 也减弱了生物膜的形成。总体而言，siamycin I 将提供一种治疗肠球菌感染的新方法。

项目成果

Development of qorum sensing inhibitors targeting enterococcal fsr system

针对肠球菌FSR系统的群体感应抑制剂的开发

• 发表时间: 2006
• 期刊: Seibutsu-kougaku Kaishi 84(10)
• 作者: Nishiguchi;K.;J.Nakayama;K.Sonomoto
• 通讯作者: K.Sonomoto

Revised model for Enterococcus faecalis fsr quorum-sensing system : small open reading frame, fsrD, encodes gelatinase biosynthesis-activating pheromone propeptide corresponding to staphylococcal AgrD

粪肠球菌 fsr 群体感应系统的修订模型：小开放阅读框，fsrD，编码与葡萄球菌 AgrD 相对应的明胶酶生物合成激活信息素前肽

• 发表时间: 2006
• 期刊: Journal of Bacteriology 188・23
• 作者: Nakayama;J.;E.Tanaka;R.Kariyama;K.Nagata;K.Nishiguchi;R.Mitsuhata;Y.Uemura;M.Tanokura;H.Kumon;K.Sonomoto;J.Nakayamaら;J.Nakayama et al.
• 通讯作者: J.Nakayama et al.

腸球菌のfsr制御系を標的としたクオーラムセンシング阻害剤開発に関する研究

针对肠球菌FSR控制系统的群体感应抑制剂的开发研究

• 发表时间: 2006
• 期刊: 生物工学 84・10
• 作者: Nishiguchi;K.;J.Nakayama;K.Sonomoto;J.Nakayamaら;西口賢三ら
• 通讯作者: 西口賢三ら

腸球菌のfsr制御系を標的としたクォーラムセンシング阻害剤開発

针对肠球菌FSR控制系统的群体感应抑制剂的开发

• 发表时间: 2006
• 期刊: 生物工学 84・10
• 作者: Nakayama;J.;E.Tanaka;R.Kariyama;K.Nagata;K.Nishiguchi;R.Mitsuhata;Y.Uemura;M.Tanokura;H.Kumon;K.Sonomoto;J.Nakayamaら;J.Nakayama et al.;西口賢三ら
• 通讯作者: 西口賢三ら

Siamycin attenuates fsr quorum sensing mediated by gelatinase biosynthesis-activating pheromone in Enterococcus faecalis

Siamycin 减弱粪肠球菌中明胶酶生物合成激活信息素介导的 fsr 群体感应

• 发表时间: 2007
• 期刊: J. Bacteriol 189
• 作者: Nakayama;J.;Tanaka;E.;Kariyama;R.;Nagata;K.;Nishiguchi;K.;Mitsuhata;R.;Uemura;Y.;Tanokura;M.;Kumon;H.;Sonomoto;K
• 通讯作者: K