Studies on Regulatory Mechanism of Salmonella Pathogenesis by Proteome Analysis

蛋白质组分析研究沙门氏菌发病机制

• 批准号: 13470058
• 负责人: YAMAMOTO Tomoko
• 金额: $ 9.15万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470058/
• 关键词: Salmonella; Proteome; Macrophage; Pathogenesis; Apoptosis; Molecular Chaperone; Lon; ClpXP; SPI1; Lonプロテアーゼ; ClpXPプロテアーゼ; 侵入性; 鞭毛; ストレス蛋白質

Intracellular pathogens, including Salmonella, which maintain long-term resistance within host phagocytes elicit a variety of genetic programs to help them adapt to the hostile environmental conditions encountered within the phagosome. To elucidate the genetic programs for Salmonella pathogenesis expressed in host after infection, a proteome analysis of proteins newly induced in Salmonella after phagocytosis by macrophages was performed. Since prominent among these programs was the heat shock response, the, role of molecular chaperones and AAA+ proteases which are members of the heat shock proteins as virulence proteins have been studied.(1)The disruption of ClpXP or Lon of serovar Typhimurium results a loss of virulence to mice. The attenuation could be due to the impaired ability to survive and replicate within macrophage cells, suggesting that ClpXP and Lon are critically important for the systemic Salmonella infection of mice. We found that the ClpXP mutant and Lon mutant persist i … More n the BALB/c mice for long periods of time without causing an overwhelming systemic infection, suggesting a possible candidate of Salmonella live vaccine. We therefore examined whether oral immunization with the lon mutant or clpXP mutant protects mice against subsequent oral challenge with virulent serovar Typhimurium. The results suggested that a single oral immunization of the lon and clpXP mutants should be effective to protect against the colonization of wild-type serovar Typhimurium within the intestinal tract.(2)We demonstrated that depletion of ATP-dependent Lon protease in serovar Typhimurium induces rapid and massive apoptosis in macrophages by a mechanism involving both caspases-1 and -3. This excessive induction of apoptosis was abrogated by disruption of invF, which is required for the expression of the Salmonella pathogenicity island 1(SPI1) genes. Expression of hilA, a central regulator of SPI1 transcription, was repressed in the macrophages after phagocytosis, but this gene was continuously expressed when the Lon mutant grew within the macrophages, so the SPI1 proteins accumulated. Thus, the increase in macrophage apoptosis induced by the Lon mutant could be due to continued expression of SPI1 genes under conditions where they are normally repressed. Once Salmonella has established a systemic infection, excess apoptosis of macrophages cells upon which the organism is reliant would be detrimental to the pathogen. Therefore, the Lon protease may be required to suppress apoptosis sufficiently to allow time for the bacterium to replicate, escape, and invade new macrophages.(3)The DnaK/DnaJ-depleted mutant lost the ability to causes a lethal systemic disease in mice. Macrophage-survival assays revealed that the mutant could not survive or proliferate at all within macrophages. Of further interest are the findings that the mutant could neither invade cultured epithelial cells nor secrete any of the invasion proteins encoded by Salmonella pathogenicity island 1. Less

包括沙门氏菌在内的细胞内病原体，这些病原体在宿主吞噬细胞内保持长期抵抗会引起各种遗传程序，以帮助它们适应吞噬体中遇到的敌对环境条件。为了阐明感染后宿主在宿主中表达的沙门氏菌发病机理的遗传程序，对巨噬细胞吞噬后吞噬作用后，对蛋白质新诱导的蛋白质组分析。由于这些程序中突出的是热休克响应，因此，分子链烷和AAA+蛋白的作用是热休克蛋白的成员，因为病毒蛋白已经研究了。衰减可能是由于在巨噬细胞中生存和复制的能力受损，这表明Clpxp和LON对于小鼠的全身沙门氏菌感染至关重要。我们发现，Clpxp突变体和LON突变体持续存在，而不是长时间的BALB/C小鼠，而不会引起压倒性的全身感染，这表明可能是沙门氏菌活疫苗的候选者。因此，我们检查了用LON突变体或ClpxP突变体口服免疫抑制是否可以保护小鼠免受随后的口服毒种typhimurium的口服挑战。 The results suggested that a single oral immunosuppression of the lon and clpXP mutants should be Effective to protect against the colonization of wild-type serovar Typhimurium within the intestinal tract.(2)We demonstrated that depletion of ATP-dependent Lon protein in serovar Typhimurium induces rapid and massive apoptosis in macrophages by a mechanism involving both caspases-1 and -3。通过破坏Invf，这种过多的凋亡诱导被废除了，这是沙门氏菌致病岛1（SPI1）基因表达所必需的。希拉（Spi1转录的中心调节剂）的表达在吞噬吞噬作用后被抑制在巨噬细胞中，但是当lon突变体在巨噬细胞内生长时，该基因被连续表达，因此SPI1蛋白积累。这，lon突变体引起的巨噬细胞凋亡的增加可能是由于在通常再现的条件下SPI1基因的持续表达。一旦沙门氏菌建立了全身感染，涉及生物体的巨噬细胞细胞的凋亡将不利于病原体。因此，可能需要LON蛋白酶充分抑制凋亡，以使细菌复制，逃脱和入侵新的巨噬细胞。巨噬细胞生存的阿萨斯（Assas）表明，突变体在巨噬细胞中根本无法生存或增殖。进一步感兴趣的是，突变体无法侵入培养的上皮细胞，也不能秘密秘密任何由沙门氏菌致病性岛编码的任何入侵蛋白1。

项目成果

Tomoyasu, T., T.Ohkishi, Y.Ukyo, A.Tokumitsu, A.Takaya, M.Suzuki, K.Sekiya, H.Matsui, K.Kutsukake, T.Yamamoto: "The ClpXP ATP-dependent protease regulates flagellum synthesis in Salmonella enterica serovar Typhimurium"J.Bacteriol.. 184(3). 645-653 (2002)

Tomoyasu, T., T.Ohkishi, Y.Ukyo, A.Tokumitsu, A.Takaya, M.Suzuki, K.Sekiya, H.Matsui, K.Kutsukake, T.Yamamoto：“ClpXP ATP 依赖性蛋白酶调节鞭毛

Takaya A., et al.: "Lon, a stress-induced ATP-dependent protease is critically important for the systemic Salmonella infection of mice"Infection and Immunity. 71. 690-696 (2003)

Takaya A. 等人：“Lon，一种应激诱导的 ATP 依赖性蛋白酶对于小鼠的全身性沙门氏菌感染至关重要”感染和免疫。

The ATP-dependent Lon protease of Salmonella enterica serovar Typhimurium regulates invasion and expression of genes carried on Salmonella pathogenicity island 1

• DOI: 10.1128/jb.184.1.224-232.2002
• 发表时间: 2002-01-01
• 期刊: JOURNAL OF BACTERIOLOGY
• 影响因子: 3.2
• 作者: Takaya, A;Tomoyasu, T;Yamamoto, T
• 通讯作者: Yamamoto, T

Takaya A., et al.: "The ATP-dependent Lon protease of Salmonella enterica serovar Typhimurium regulates invasion and expression of genes encoded on Salmonella pathogenicity island"Journal of Bacteriology. 184. 224-232 (2002)

Takaya A. 等人：“鼠伤寒沙门氏菌肠炎血清型的 ATP 依赖性 Lon 蛋白酶调节沙门氏菌致病岛编码基因的侵袭和表达”《细菌学杂志》。

医学大辞典

医学词典

• 发表时间: 2006
• 作者: 末広 寛;Heather Skirton;村上 京子;塚原正人;Skirton H;飯野英親;飯野英親;塚原正人;辻野久美子;松本浩;飯野英親;飯野英親;川内美穂子;斉藤知子;村上京子;末広 寛;辻野久美子;Matsumoto Y;飯野英親;村上京子;飯野英親;中込さと子;塚原正人;塚原正人;塚原 正人;飯野 英親;辻野 久美子;村上 京子;飯野 英親;辻野 久美子;石田喬士;相澤忠範
• 通讯作者: 相澤忠範