Targeting of a chlamydial protease impedes intracellular bacterial growth.
Targeting of a chlamydial protease impedes intracellular bacterial growth.
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DOI：
10.1371/journal.ppat.1002283
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发表时间：
2011-09
期刊：
影响因子：
6.7
通讯作者：
Heuer D
中科院分区：
文献类型：
--
作者：
Christian JG;Heymann J;Paschen SA;Vier J;Schauenburg L;Rupp J;Meyer TF;Häcker G;Heuer D
衣原体是在胞质液泡中繁殖的专性细胞内细菌。最近的研究表明，衣原体的生长会诱导高尔基体 (GA) 破碎成微型堆栈，这有助于将宿主脂质获取到不断生长的内含物中。 GA 碎片是由感染相关的完整 GA 蛋白 golgin-84 的裂解引起的。肽抑制剂 WEHD-fmk 可以阻断 Golgin-84 裂解、GA 断裂和沙眼衣原体生长。在这里，我们将细菌蛋白酶衣原体蛋白酶样活性因子 (CPAF) 确定为介导 golgin-84 裂解的因子，并作为 WEHD-fmk 抑制的靶标。 WEHD-fmk 阻断了沙眼衣原体和肺炎衣原体感染期间 golgin-84 的裂解以及已知 CPAF 靶标的裂解。当活性 CPAF 在未感染细胞和无细胞系统中表达时，观察到相同的效果。未感染细胞中活性 CPAF 的异位表达足以导致 GA 碎片化。 GA 片段化需要 CPAF 活性下游的小型 GTPases Rab6 和 Rab11。这些结果将 CPAF 定义为衣原体复制所必需的第一种蛋白质。我们认为这种作用使 CPAF 成为潜在的抗感染治疗靶点。衣原体是仅在宿主（例如人类）细胞内复制的细菌，是人类疾病（特别是性传播疾病）的常见病原体。衣原体生活在细胞内的液泡中，周围有脂质膜，必须从宿主细胞获取营养和其他因子才能复制和液泡生长。最近的结果表明，为此，衣原体依赖于其诱导高尔基体（一种对细胞内运输物质进行分类的细胞结构）的单个蛋白质（称为 golgin-84）丢失的能力。在这项工作中，我们发现衣原体利用其蛋白质切割酶 CPAF（由衣原体产生并从液泡转运到细胞中）来做到这一点。 CPAF 裂解 golgin-84，从而诱导高尔基体发生变化，这些变化与衣原体获取某些细胞物质有关。我们进一步表明，最近发现的一种合成抑制剂可以通过抑制 CPAF 来阻止衣原体生长。因此，CPAF 似乎对于衣原体生长是必要的，并且阻断 CPAF 可能是针对衣原体感染的治疗策略。
Chlamydiae are obligate intracellular bacteria that propagate in a cytosolic vacuole. Recent work has shown that growth of Chlamydia induces the fragmentation of the Golgi apparatus (GA) into ministacks, which facilitates the acquisition of host lipids into the growing inclusion. GA fragmentation results from infection-associated cleavage of the integral GA protein, golgin-84. Golgin-84-cleavage, GA fragmentation and growth of Chlamydia trachomatis can be blocked by the peptide inhibitor WEHD-fmk. Here we identify the bacterial protease chlamydial protease-like activity factor (CPAF) as the factor mediating cleavage of golgin-84 and as the target of WEHD-fmk-inhibition. WEHD-fmk blocked cleavage of golgin-84 as well as cleavage of known CPAF targets during infection with C. trachomatis and C. pneumoniae. The same effect was seen when active CPAF was expressed in non-infected cells and in a cell-free system. Ectopic expression of active CPAF in non-infected cells was sufficient for GA fragmentation. GA fragmentation required the small GTPases Rab6 and Rab11 downstream of CPAF-activity. These results define CPAF as the first protein that is essential for replication of Chlamydia. We suggest that this role makes CPAF a potential anti-infective therapeutic target. Chlamydiae are bacteria that replicate only inside host (for instance human) cells and that are frequent agents of human disease, in particular sexually transmitted disease. Chlamydia lives in a vacuole inside the cell, surrounded by a lipid membrane, and must acquire nutrients and other factors from the host cell for its replication and for the growth of the vacuole. Recent results show that for this, Chlamydia relies on its ability to induce the loss of an individual protein of the Golgi apparatus (a cellular structure that sorts materials for transport in the cell) called golgin-84. In this work we find that Chlamydia does this using its protein-cleaving enzyme CPAF (which is made by Chlamydia and transported from the vacuole into the cell). CPAF cleaves golgin-84 and thereby induces changes in the Golgi apparatus that are linked to the acquisition of some cellular material by Chlamydia. We further show that a synthetic inhibitor, which was recently found to block chlamydial growth, does that by inhibiting CPAF. CPAF therefore seems necessary for chlamydial growth and blocking CPAF may be a therapeutic strategy against infections with Chlamydia.
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影响因子：
3.1
作者：
Dong, F;Su, H;Zhong, GM
通讯作者：
Zhong, GM
影响因子：
168.9
作者：
Gerbase, AC;Rowley, JT;Mertens, TE
通讯作者：
Mertens, TE
影响因子：
56.9
作者：
Horn, M;Collingro, A;Wagner, M
通讯作者：
Wagner, M
DOI：
10.1073/pnas.92.11.4877
发表时间：
1995-05-23
影响因子：
11.1
作者：
HACKSTADT, T;SCIDMORE, MA;ROCKEY, DD
通讯作者：
ROCKEY, DD
影响因子：
3.4
作者：
Heuer, D;Brinkmann, V;Szczepek, AJ
通讯作者：
Szczepek, AJ