Listeria monocytogenes is an important zoonotic foodborne pathogen that can tolerate a number of environmental stresses. Rsb R, an upstream regulator of the sigma B(Sig B) factor, is thought to sense environmental cha...Listeria monocytogenes is an important zoonotic foodborne pathogen that can tolerate a number of environmental stresses. Rsb R, an upstream regulator of the sigma B(Sig B) factor, is thought to sense environmental challenges and trigger the SigB pathway. In Bacillus subtilis, two phosphorylation sites in RsbR are involved in activating the SigB pathway and a feedback mechanism, respectively. In this study, the role of RsbR in L. monocytogenes under mild and severe stresses was investigated. Strains with genetic deletion(Δrsb R), complementation(C-Δrsb R), and phosphorylation site mutations in the rsbR(RsbR-T175 A, RsbR-T209 A, and RsbR-T175 A-T209 A) were constructed to evaluate the roles of these RsbR sequences in listerial growth and survival. SigB was examined at the transcriptional and translational levels. Deletion of rsb R reduced listerial growxth and survival in response to acidic stress. Substitution of the phosphorylation residue RsbR-T175 A disabled RsbR complementation, while RsbR-T209 A significantly upregulated SigB expression and listerial survival. Our results provide clear evidence that two phosphorylation sites of Rsb R are functional in L. monocytogenes under acidic conditions, similar to the situation in B. subtilis.展开更多
目的探讨运用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)技术对金黄色葡萄球菌小菌落变异型的特征性蛋白指纹图谱进行深度研究的价值。方法分别...目的探讨运用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)技术对金黄色葡萄球菌小菌落变异型的特征性蛋白指纹图谱进行深度研究的价值。方法分别采用传统生化鉴定、16S r RNA测序技术和MALDI-TOF MS技术对1株小菌落变异型金黄色葡萄球菌进行鉴定,通过质谱鉴定系统VITEK MS的质谱科研模式获取金黄色葡萄球菌小菌落突变株(small colony variants,SCV)的质谱图,分析代表金黄色葡萄球菌agr和sig B系统的m/z表达强度,并用SARAMIS软件构建基于质谱峰谱特征的系统发育树。结果使用MALDI-TOF MS技术鉴定该菌株为金黄色葡萄球菌,与传统生化鉴定、16S rRNA测序技术的鉴定结果一致。其菌落形态符合典型SCV菌株的特征,预示生物膜形成的PSMα3(m/z 2634.4)、PSMα1(m/z 2286.8)和PSMα4(m/z 2198.6)的表达强度分别为100%、76.1%和32.3%,反映急性早期感染状态的agr调控系统的delta毒素强度只有10%;反映慢性、复发性感染的sigB调控系统的峰谱相对强度均<25%,该菌株和科研菌种库SCV的系统发育树近似度>70%。结论 MALDI-TOF MS技术不仅能够快速准确鉴定不典型金黄色葡萄球菌,且生成的质谱图能够初步提示菌株在宿主体内的感染状态,表明该技术具有极大的应用前景。展开更多
基金Project supported by the National Natural Science Foundation of China(Nos.31702250 and 31600292)the Open Fund Project by Key Laboratory of Animal Preventive Medicine of Zhejiang Province(No.ZPKLPVM2017KFKT001),China
文摘Listeria monocytogenes is an important zoonotic foodborne pathogen that can tolerate a number of environmental stresses. Rsb R, an upstream regulator of the sigma B(Sig B) factor, is thought to sense environmental challenges and trigger the SigB pathway. In Bacillus subtilis, two phosphorylation sites in RsbR are involved in activating the SigB pathway and a feedback mechanism, respectively. In this study, the role of RsbR in L. monocytogenes under mild and severe stresses was investigated. Strains with genetic deletion(Δrsb R), complementation(C-Δrsb R), and phosphorylation site mutations in the rsbR(RsbR-T175 A, RsbR-T209 A, and RsbR-T175 A-T209 A) were constructed to evaluate the roles of these RsbR sequences in listerial growth and survival. SigB was examined at the transcriptional and translational levels. Deletion of rsb R reduced listerial growxth and survival in response to acidic stress. Substitution of the phosphorylation residue RsbR-T175 A disabled RsbR complementation, while RsbR-T209 A significantly upregulated SigB expression and listerial survival. Our results provide clear evidence that two phosphorylation sites of Rsb R are functional in L. monocytogenes under acidic conditions, similar to the situation in B. subtilis.
