Control of type III protein secretion using a minimal genetic system.
Control of type III protein secretion using a minimal genetic system.
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DOI：
10.1038/ncomms14737
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发表时间：
2017-05-09
影响因子：
16.6
通讯作者：
Voigt CA
中科院分区：
文献类型：
--
作者：
Song M;Sukovich DJ;Ciccarelli L;Mayr J;Fernandez-Rodriguez J;Mirsky EA;Tucker AC;Gordon DB;Marlovits TC;Voigt CA
革兰氏阴性细菌使用 III 型分泌系统 (T3SS) 分泌蛋白质，该系统起到针状分子机器的作用。参与 T3SS 构建的许多蛋白质由于其在发病机制和运动中的作用而受到严格调控。在这里，从 35 kb 沙门氏菌致病性岛 1 (SPI-1) 开始，我们消除了内部调节，并通过删除或重新编码基因、打乱基因顺序以及用合成遗传部分替换所有非编码 DNA 来简化遗传学。这个过程产生了一个 16kb 的簇，它与 SPI-1 没有序列同一性、规则或组织原则。构建这个简化的系统导致内部起始位点 (SpaO) 和小 RNA (InvR) 的重要作用的发现。此外，它可以使用合成调节电路进行控制，包括在 SPI-1 抑制条件下。这项工作揭示了 T3SS 组装令人难以置信的转录后稳健性，并有助于其作为生物技术工具的控制。 III型分泌系统是一个受到严格监管控制的针状分子机器。在此，作者通过解构和重建野生型沙门氏菌致病岛1，构建了合成的III型分泌系统基因簇。
Gram-negative bacteria secrete proteins using a type III secretion system (T3SS), which functions as a needle-like molecular machine. The many proteins involved in T3SS construction are tightly regulated due to its role in pathogenesis and motility. Here, starting with the 35 kb Salmonella pathogenicity island 1 (SPI-1), we eliminated internal regulation and simplified the genetics by removing or recoding genes, scrambling gene order and replacing all non-coding DNA with synthetic genetic parts. This process results in a 16 kb cluster that shares no sequence identity, regulation or organizational principles with SPI-1. Building this simplified system led to the discovery of essential roles for an internal start site (SpaO) and small RNA (InvR). Further, it can be controlled using synthetic regulatory circuits, including under SPI-1 repressing conditions. This work reveals an incredible post-transcriptional robustness in T3SS assembly and aids its control as a tool in biotechnology. The type III secretion system is a needle-like molecular machine under tight regulatory control. Here the authors construct a synthetic type III secretion system gene cluster by deconstructing and rebuilding the wild-type Salmonella pathogenicity island 1.