Targeted Disruption of Chlamydia trachomatis Invasion by in Trans Expression of Dominant Negative Tarp Effectors.
Targeted Disruption of Chlamydia trachomatis Invasion by in Trans Expression of Dominant Negative Tarp Effectors.
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DOI：
10.3389/fcimb.2016.00084
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发表时间：
2016
影响因子：
5.7
通讯作者：
Jewett TJ
中科院分区：
文献类型：
--
作者：
Parrett CJ;Lenoci RV;Nguyen B;Russell L;Jewett TJ
III 型分泌效应蛋白 Tarp 部分促进了沙眼衣原体侵入真核宿主细胞。 Tarp 在衣原体进入宿主细胞中的作用得到了分子方法的支持，这些方法检查了异源系统中表达的重组 Tarp 或 Tarp 效应子。研究 Tarp 对衣原体入侵宿主细胞的作用的一个主要限制是先前缺乏衣原体遗传工具。基于我们对 Tarp 结构域结构和功能的了解以及在沙眼衣原体中引入遗传方法，我们假设通过引入显性失活突变等位基因可以在体内破坏 Tarp 功能。我们提供的证据表明，转化的沙眼衣原体产生了标记有 Tarp 的表位，该表位在入侵过程中被分泌到宿主细胞中。我们检查了域特异性 Tarp 突变对衣原体入侵和生长的影响，并证明携带缺乏肌动蛋白结合域或磷酸化域的工程 Tarp 突变体的沙眼衣原体克隆降低了对宿主细胞的入侵水平。这些数据为 Tarp 在沙眼衣原体发病机制中的关键作用提供了第一个体内证据，并表明可以通过引入工程化的显性失活三型效应子来减弱衣原体对宿主细胞的入侵。
Chlamydia trachomatis invasion of eukaryotic host cells is facilitated, in part, by the type III secreted effector protein, Tarp. The role of Tarp in chlamydiae entry of host cells is supported by molecular approaches that examined recombinant Tarp or Tarp effectors expressed within heterologous systems. A major limitation in the ability to study the contribution of Tarp to chlamydial invasion of host cells was the prior absence of genetic tools for chlamydiae. Based on our knowledge of Tarp domain structure and function along with the introduction of genetic approaches in C. trachomatis, we hypothesized that Tarp function could be disrupted in vivo by the introduction of dominant negative mutant alleles. We provide evidence that transformed C. trachomatis produced epitope tagged Tarp, which was secreted into the host cell during invasion. We examined the effects of domain specific Tarp mutations on chlamydial invasion and growth and demonstrate that C. trachomatis clones harboring engineered Tarp mutants lacking either the actin binding domain or the phosphorylation domain had reduced levels of invasion into host cells. These data provide the first in vivo evidence for the critical role of Tarp in C. trachomatis pathogenesis and indicate that chlamydial invasion of host cells can be attenuated via the introduction of engineered dominant negative type three effectors.