Localization of Chlamydia trachomatis hypothetical protein CT311 in host cell cytoplasm.
Localization of Chlamydia trachomatis hypothetical protein CT311 in host cell cytoplasm.
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沙眼衣原体的定位假设蛋白CT311在宿主细胞细胞质中的定位。
DOI：
10.1016/j.micpath.2011.05.002
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发表时间：
2011-09
影响因子：
3.8
通讯作者：
Zhong G
中科院分区：
文献类型：
--
作者：
Lei L;Qi M;Budrys N;Schenken R;Zhong G
在沙眼衣原体感染细胞的衣原体包涵体内部和外部均检测到衣原体特异性假设蛋白 CT311。额外包涵体 CT311 分子以与 CPAF（一种已知的衣原体分泌蛋白酶）类似的模式分布在宿主细胞的细胞质中。 CT311的检测是特异性的，因为抗CT311抗体标记仅通过CT311的吸收而被去除，而CPAF融合蛋白则未被去除。此外，抗CT311和抗CPAF抗体均仅检测其相应的内源蛋白，而不会与沙眼衣原体感染细胞的全细胞裂解物中的彼此或任何其他抗原发生交叉反应。虽然CT311和CPAF蛋白在感染后12小时首次被检测到，但CT311定位到宿主细胞胞质中的时间被延迟到24小时，而CPAF分泌到宿主细胞胞质中的情况在感染后18小时就已经很明显了。 CT311 的宿主细胞胞浆定位在人原代细胞中得到进一步证实。预计 CT311 含有 N 端分泌信号序列，并且 CT311 信号序列在异源测定系统中指导 PhoA 分泌到细菌周质区域，这表明 sec 依赖性途径可能在 CT311 分泌到宿主细胞胞浆中的过程中发挥作用。这一假设得到了观察结果的进一步支持，即衣原体感染细胞中 CT311 的分泌被已知抑制信号肽酶 I 的 C16 化合物阻断。这些发现为进一步了解沙眼衣原体致病机制提供了重要的分子信息。
The chlamydia-specific hypothetical protein CT311 was detected both inside and outside of the chlamydial inclusions in C. trachomatis-infected cells. The extra-inclusion CT311 molecules were distributed in the host cell cytoplasm with a pattern similar to that of CPAF, a known Chlamydia-secreted protease. The detection of CT311 was specific since the anti-CT311 antibody labeling was only removed by absorption with CT311 but not CPAF fusion proteins. In addition, both anti-CT311 and anti-CPAF antibodies only detected their corresponding endogenous proteins without cross-reacting with each other or any other antigens in the whole cell lysates of C. trachomatis-infected cells. Although both CT311 and CPAF proteins were first detected 12h after infection, localization of CT311 into host cell cytosol was delayed until 24h while CPAF secretion into host cell cytosol was already obvious by 18h after infection. The host cell cytosolic localization of CT311 was further confirmed in human primary cells. CT311 was predicted to contain a N-terminal secretion signal sequence and the CT311 signal sequence directed secretion of PhoA into bacterial periplasmic region in a heterologous assay system, suggesting that a sec-dependent pathway may play a role in the secretion of CT311 into host cell cytosol. This hypothesis is further supported by the observation that secretion of CT311 in Chlamydia-infected cells was blocked by a C16 compound known to inhibit signal peptidase I. These findings have provided important molecular information for further understanding the C. trachomatis pathogenic mechanisms.
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