Proteomic and functional analysis of the suite of Ysp proteins exported by the Ysa type III secretion system of Yersinia enterocolitica Biovar 1B
Proteomic and functional analysis of the suite of Ysp proteins exported by the Ysa type III secretion system of Yersinia enterocolitica Biovar 1B
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DOI：
10.1111/j.1365-2958.2005.04973.x
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发表时间：
2006-01-01
影响因子：
3.6
通讯作者：
Young, GM
中科院分区：
文献类型：
--
作者：
Matsumoto, H;Young, GM
小肠结肠炎耶尔森氏菌生物型1B的完全毒力需要两个不同且关系较远的接触依赖性III型分泌（T3 S）系统。质粒编码的Ysc T3 S系统对于感染的全身性阶段是必需的，并且其易位的Yop效应蛋白已被广泛研究。染色体编码的Ysa T3 S系统有助于感染的胃肠道阶段，但它易位到宿主细胞中的Ysp效应蛋白套件仍然不清楚。使用基于蛋白质组学的方法，分析Ysa T3 S系统，揭示了一组复杂的15个分泌的Ysp蛋白。这些蛋白质中的七种先前被描述（YspA、YspB、YspC、YspD、YopE、YopN和YopP）。这些Ysps中的八种（YspK、YspI、YspE、YspF、YspP、YspY、YspN和YspL）先前未被表征。一些新的Ysps与其他毒力因子同源，包括YspP与耶尔森氏菌蛋白酪氨酸磷酸酶YopH相似，YspK与志贺氏菌丝氨酸/苏氨酸激酶OspG相似。纯化的六组氨酸标记的YspK和YspP的生化分析确定，这些蛋白质分别具有激酶和磷酸酶活性。用Y.表达Ysp-CyaA嵌合蛋白的小肠结肠炎菌菌株导致六种Ysps（YspK、YspI、YspE、YspF、YspP和YspL）的cAMP胞质水平的Ysa T3 S系统依赖性增加，而不是另外两种（YspY和YspN）。然而，YspN，所需的效应蛋白易位到真核细胞的Ysa T3 S系统。在BALB/c小鼠中的竞争测定显示，单个Ysp产生缺陷的突变体影响胃肠道组织的定殖。总的来说，这项研究的结果支持了这样的假设，即Ysa T3 S系统将一套复杂的效应蛋白靶向宿主细胞，以影响感染的结果。由Ysa T3 S系统递送的效应子套件的鉴定揭示了宿主细胞信号传导途径是几种Ysp效应子的可能靶标。
Full virulence of Yersinia enterocolitica Biovar 1B requires two distinct and distantly related contact-dependent type III secretion (T3S) systems. The plasmid-encoded Ysc T3S system is essential for systemic stages of infection and the Yop effector proteins it translocates have been extensively studied. The chromosome-encoded Ysa T3S system contributes to gastrointestinal stages of infection, but the suite of Ysp effectors proteins it translocates into host cells remains obscure. Using a proteomics-based approach, the Ysa T3S system was analysed revealing a complex set of 15 secreted Ysp proteins. Seven of these proteins were previously described (YspA, YspB, YspC, YspD, YopE, YopN and YopP). Eight of these Ysps (YspK, YspI, YspE, YspF, YspP, YspY, YspN and YspL) had not previously been characterized. Several of the new Ysps are homologous to other virulence factors, including YspP with similarity to the Yersinia protein tyrosine phosphatase YopH and YspK with similarity to the Shigella serine/threonine kinase OspG. Biochemical analysis of purified hexa-histidine tagged YspK and YspP established that these proteins have kinase and phosphatase activity respectively. Infection of eukaryotic cells with Y. enterocolitica strains expressing a Ysp-CyaA chimeric protein resulted in Ysa T3S system-dependent increases in cytosolic levels of cAMP for six Ysps (YspK, YspI, YspE, YspF, YspP and YspL), but not two others (YspY and YspN). YspN, however, was required for translocation of effector proteins into eukaryotic cells by the Ysa T3S system. Competition assays in BALB/c mice revealed that mutants defective for the production of an individual Ysp are affected for colonization of gastrointestinal tissues. Collectively, the results of this study support the hypothesis that the Ysa T3S system targets a complex suite of effector proteins into host cells to affect the outcome of an infection. Identification of the suite of effectors delivered by the Ysa T3S system reveals that host cell signalling pathways are the probable targets of several Ysp effectors.