Chlamydia pneumoniae inclusion membrane protein cpn0585 interacts with multiple rab GTPases
Chlamydia pneumoniae inclusion membrane protein cpn0585 interacts with multiple rab GTPases
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DOI：
10.1128/iai.01020-07
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发表时间：
2007-12-01
影响因子：
3.1
通讯作者：
Wizel, Benjamin
中科院分区：
文献类型：
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作者：
Cortes, Claudio;Rzomp, Mmberly A.;Wizel, Benjamin
衣原体是细胞内细菌，在称为包涵体的膜结合空泡内发育。为了确保包涵体是一个安全的小生境，以chiamydial复制，衣原体利用了一些宿主细胞的过程，包括膜运输途径。最近，发现几种Rab GTP酶与各种衣原体的包涵体有关。在这里，我们报告Cpn 0585，肺炎衣原体包涵体膜蛋白（公司），与多个Rab GTP酶相互作用。酵母双杂交实验的结果表明，Cpn 0585的氨基末端截短形式（Cpn 0585（102-651））与Rab 1，Rab 10和Rab 11相互作用，但不与Rab 4或Rab 6相互作用。Cpn 0585-Rab GTP酶相互作用是直接的和GTP依赖性的，如使用天然和重组Cpn 0585的谷胱甘肽S-转移酶下拉测定所示。In C.在用增强型绿色荧光蛋白（EGFP）标记的Rab GTP酶转染的肺炎链球菌感染的HEp-2细胞中，Cpn 0585在包涵体膜上的共定位对于EGFP-Rab 1和EGFPRab 10是部分的，但是对于野生型EGFP-Rab 11 A和组成型活性GTP酶缺陷型EGFPRab 11 AQ 70 L是广泛的。Cpn 0585与EGFP-Rab 11 AQ 70 L在感染后2 h即发生共定位。在交付到现场C。在肺炎球菌感染的细胞中，Cpn 0585（628-651）特异性抗体结合到包涵体膜，证明Cpn 0585的Rab GTP酶相互作用结构域面向宿主细胞胞质溶胶。Cpn 0585（102-651）的异位表达部分抑制了C. pneumoniae包涵体。而在表达EGFP-Rab 11 AQ 70 L的HEp-2细胞中则没有。总的来说，这些数据表明Cpn 0585参与了Rab GTP酶向包涵体膜的募集，干扰这一功能可能会对C. pneumoniae包涵体进行衣原体复制。
Chlamydiae are intracellular bacteria that develop within a membrane-bound vacuole called an inclusion. To ensure that the inclusion is a safe niche for chiamydial replication, chlamydiae exploit a number of host cell processes, including membrane-trafficking pathways. Recently, several Rab GTPases were found to associate with the inclusions of various chlamydial species. Here we report that Cpn0585, a Chlamydia pneumoniae inclusion membrane protein (Inc), interacts with multiple Rab GTPases. The results from yeast two-hybrid experiments revealed that an amino-terminally truncated form of Cpn0585 (Cpn0585(102-651)) interacts with Rab1, Rab10, and Rab11 but not with Rab4 or Rab6. Cpn0585-Rab GTPase interactions are direct and GTP dependent as shown in glutathione S-transferase pull-down assays using native and recombinant Cpn0585. In C. pneumoniae-infected HEp-2 cells transfected with enhanced green fluorescent protein (EGFP)-tagged Rab GTPases, the colocalization with Cpn0585 at the inclusion membrane was partial for EGFP-Rab1 and EGFPRab10, but extensive for wild-type EGFP-Rab11A and the constitutively active GTPase-deficient EGFPRab11AQ70L. Moreover, Cpn0585 colocallized with EGFP-Rab11AQ70L as early as 2 h postinfection. Upon delivery into live C. pneumoniae-infected cells, Cpn0585(628-651)-specific antibodies bound to the inclusion membrane, demonstrating that the Rab GTPase-interacting domain of Cpn0585 faces the host cell cytosol. Finally, ectopic expression of Cpn0585(102-651) partially inhibited the development of C. pneumoniae inclusions in EGFP. but not in EGFP-Rab11AQ70L-expressing HEp-2 cells. Collectively, these data suggest that Cpn0585 is involved in the recruitment of Rab GTPases to the inclusion membrane and that interfering with this function may adversely impact the fitness of the C. pneumoniae inclusion for chlamydial replication.