Cell-selective proteomics reveal novel effectors secreted by an obligate intracellular bacterial pathogen.
Cell-selective proteomics reveal novel effectors secreted by an obligate intracellular bacterial pathogen.
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细胞选择性蛋白质组学揭示了专性细胞内细菌病原体分泌的新型效应子。
DOI：
10.1101/2023.11.17.567466
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Lamason,RebeccaL
中科院分区：
文献类型：
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作者：
Sanderlin,AllenG;Margolis,HannahK;Meyer,AbigailF;Lamason,RebeccaL
病原菌分泌蛋白质效应子劫持宿主机器并重塑其感染生态位。立克次体是专性细胞内细菌，可导致危及生命的疾病，但它们对宿主细胞的绝对依赖阻碍了立克次体效应子及其宿主靶点的发现。我们在R.本发明涉及一种用于介导parkeriinfection以选择性地标记、分离和鉴定递送到宿主细胞中的效应物的方法。作为BONCAT在专性细胞内细菌中的第一次使用，我们的筛选使该属实验验证的效应子的数量增加了一倍以上。我们鉴定的7种新的分泌型立克次体因子（Srfs）包括定位于宿主细胞质、线粒体和ER的功能未知的立克次体特异性蛋白。我们进一步表明，一个这样的效应，SrfD，与主机Sec 61的translocon相互作用。总而言之，我们的工作揭示了一组不同的先前未知的立克次体效应子，并奠定了基础，更深入地探索宿主-病原体界面。
Pathogenic bacteria secrete protein effectors to hijack host machinery and remodel their infectious niche.Rickettsiaspp. are obligate intracellular bacteria that can cause life-threatening disease, but their absolute dependence on the host cell has impeded discovery of rickettsial effectors and their host targets. We implemented bioorthogonal non-canonical amino acid tagging (BONCAT) duringR. parkeriinfection to selectively label, isolate, and identify effectors delivered into the host cell. As the first use of BONCAT in an obligate intracellular bacterium, our screen more than doubles the number of experimentally validated effectors for the genus. The seven novel secreted rickettsial factors (Srfs) we identified includeRickettsia-specific proteins of unknown function that localize to the host cytoplasm, mitochondria, and ER. We further show that one such effector, SrfD, interacts with the host Sec61 translocon. Altogether, our work uncovers a diverse set of previously uncharacterized rickettsial effectors and lays the foundation for a deeper exploration of the host-pathogen interface.