假定调控蛋白STM14_3514可降低鼠伤寒沙门菌对上皮细胞的侵袭力  

Putative regulatory protein STM14_3514 decreases Salmonella Typhimurium invasion of epithelial cells

作　　者：蒋玲艳[1,2,3] 周启星[1] 王培胜[2,3] 江小涵 冯露[2,3] 

机构地区：[1]南开大学环境科学与工程学院,天津300350 [2]南开大学泰达生物技术研究院,天津300457 [3]天津市微生物功能基因组学重点实验室,天津300457

出　　处：《微生物学报》2017年第4期500-512,共13页Acta Microbiologica Sinica

基　　金：中国博士后科学基金(2016M591381);国家国际科技合作专项(2012DFG31680)~~

摘　　要：【目的】研究鼠伤寒沙门菌致病岛1(SPI-1)内部的假定调控蛋白STM14_3514的功能及其作用机制。【方法】以鼠伤寒沙门菌模式菌株ATCC 14028为亲本株,构建了STM14_3514基因的缺失突变体及互补菌株,通过小鼠实验、细胞侵袭实验、Western blot及实时荧光定量PCR(q RT-PCR)等实验技术,深入研究了STM14_3514基因对鼠伤寒沙门菌致病过程的影响。【结果】STM14_3514突变提高了细菌对小鼠的致病能力,突变体在小鼠肠道、肝和脾中的定殖能力均增强;细胞实验揭示,突变体致病力提升主要由于STM14_3514突变能显著增强细菌对上皮细胞的侵袭力(>2倍,P<0.05)。q RT-PCR及Western blot分析表明,STM14_3514显著抑制SPI-1内部主要调控因子hil A及侵袭相关基因的表达。此外,STM14_3514对hil A的抑制由Hil C介导。【结论】STM14_3514是鼠伤寒沙门菌SPI-1内部的负调控因子,能通过Hil C抑制hil A及SPI-1其他入侵基因的表达,该基因的生物学意义可能与细菌进入细胞后对SPI-1的负调控相关。[Objective] To study the function and mechanism of STM14_3514 gene that encoded in Salmonella pathogenicity island（SPI）-1 of Salmonella enterica serovar Typhimurium strain ATCC 14028. [Methods] We constructed STM14_3514 mutant strain and a complemented strain of the mutant. Through mice experiment, attachment assays, invasion assays, macrophage replication assays, Western blot, and Quantitative real-time PCR analysis（q RT-PCR）, we compared the virulence of the mutant strain to that of the wild-type 14028. [Results] STM14_3514 mutant shows increased virulence to mice, and the bacterial number of STM14_3514 mutant in liver, spleen, and ileum was more abundant than that of the wild-type strain. The increased virulence of STM14_3514 mutant is caused by its elevated invasion ability to epithelial cells（〉2-fold and P〈0.05）. q RT-PCR and Western blot results show that STM14_3514 reduced the expression of Hil A and another SPI-1invasion locus. Moreover, the repression of Hil A by STM14_3514 is mediated by Hil C. [Conclusion] STM14_3514 is a negative regulator in SPI-1, which can repress Hil A and SPI-1invasion locus through Hil C, and possibly contribute to the repression on SPI-1 after bacterial invasion.

关 键 词：鼠伤寒沙门菌 STM14_3514 侵袭力 Hi1A Hi1C 

分 类 号：R378[医药卫生—病原生物学]