Identification of novel type III secretion chaperone-substrate complexes of Chlamydia trachomatis.
Identification of novel type III secretion chaperone-substrate complexes of Chlamydia trachomatis.
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DOI：
10.1371/journal.pone.0056292
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Mota LJ
中科院分区：
文献类型：
--
作者：
Pais SV;Milho C;Almeida F;Mota LJ
沙眼衣原体是人类的一种专性细胞内细菌病原体，其使用III型分泌（T3 S）系统通过将效应蛋白递送到宿主细胞的胞质溶胶和细胞膜中来操纵宿主细胞。T3 S系统的功能依赖于小的细菌胞质伴侣样蛋白，其结合T3 S底物并确保其适当分泌。寻找新的C.我们首先在沙眼衣原体的基因组中寻找编码具有T3 S分子伴侣特征的蛋白质的基因。然后，我们系统地测试了候选分子伴侣和衣原体T3 S底物之间的相互作用细菌双杂交。这揭示了Slc 1（一种已知的T3 S分子伴侣）或CT 584与几种T3 S底物之间的相互作用。小肠结肠炎耶尔森氏菌蛋白表达后的免疫共沉淀和蛋白覆盖结合试验表明，Slc 1与已知T3 S底物Tarp（先前描述的Slc 1底物）、CT694和CT695的N-末端区域相互作用，CT 584与CT 082的中心区域相互作用，我们将其鉴定为C.沙眼衣原体T3 S底物。小肠结肠炎菌作为异源系统。在耶尔森氏菌中的进一步T3 S测定表明，Slc 1或CT 584分别增加分泌的Tarp、CT694和CT695或CT 082的量。CT 584的表达增加了CT 082的细菌内稳定性，而Slc 1不影响其底物的稳定性。总体而言，这表明在C.沙眼衣原体Slc 1是多种T3 S底物的伴侣，而CT 584是新鉴定的T3 S底物CT 082的伴侣。
Chlamydia trachomatis is an obligate intracellular bacterial pathogen of humans that uses a type III secretion (T3S) system to manipulate host cells through the delivery of effector proteins into their cytosol and membranes. The function of T3S systems depends on small bacterial cytosolic chaperone-like proteins, which bind T3S substrates and ensure their appropriate secretion. To find novel T3S chaperone-substrate complexes of C. trachomatis we first searched its genome for genes encoding proteins with features of T3S chaperones. We then systematically tested for interactions between candidate chaperones and chlamydial T3S substrates by bacterial two-hybrid. This revealed interactions between Slc1 (a known T3S chaperone) or CT584 and several T3S substrates. Co-immunoprecipation after protein expression in Yersinia enterocolitica and protein overlay binding assays indicated that Slc1 interacted with the N-terminal region of the known T3S substrates Tarp (a previously described substrate of Slc1), CT694, and CT695, and that CT584 interacted with a central region of CT082, which we identified as a C. trachomatis T3S substrate using Y. enterocolitica as a heterologous system. Further T3S assays in Yersinia indicated that Slc1 or CT584 increased the amount of secreted Tarp, CT694, and CT695, or CT082, respectively. Expression of CT584 increased the intra-bacterial stability of CT082, while Slc1 did not affect the stability of its substrates. Overall, this indicated that in C. trachomatis Slc1 is a chaperone of multiple T3S substrates and that CT584 is a chaperone of the newly identified T3S substrate CT082.
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影响因子：
3.6
作者：
Hower S;Wolf K;Fields KA
通讯作者：
Fields KA
影响因子：
3.6
作者：
Fields, KA;Hackstadt, T
通讯作者：
Hackstadt, T
影响因子：
16
作者：
Birtalan, SC;Phillips, RM;Ghosh, P
通讯作者：
Ghosh, P
影响因子：
6.7
作者：
Arnold R;Brandmaier S;Kleine F;Tischler P;Heinz E;Behrens S;Niinikoski A;Mewes HW;Horn M;Rattei T
通讯作者：
Rattei T
DOI：
10.1073/pnas.98.3.1160
发表时间：
2001-01-30
影响因子：
11.1
作者：
Fling, SP;Sutherland, RA;Starnbach, MN
通讯作者：
Starnbach, MN