The SycN/YscB chaperone-binding domain of YopN is required for the calcium-dependent regulation of Yop secretion by Yersinia pestis.
The SycN/YscB chaperone-binding domain of YopN is required for the calcium-dependent regulation of Yop secretion by Yersinia pestis.
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DOI：
10.3389/fcimb.2013.00001
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发表时间：
2013
影响因子：
5.7
通讯作者：
Plano GV
中科院分区：
文献类型：
--
作者：
Joseph SS;Plano GV
许多革兰氏阴性细菌病原体利用III型分泌系统（t3ss）将效应蛋白注入真核细胞。III型分泌（T3S）过程的激活在所有t3ss中都受到严格控制。在鼠疫耶尔森氏菌中，被称为耶尔森氏菌外蛋白（Yops）的效应蛋白的分泌受YopN/SycN/YscB/TyeA复合物活性的调节。YopN是一种分泌蛋白，通过n端伴侣结合域（CBD）与SycN/YscB伴侣相互作用，通过c端tya - a结合域（TBD）与TyeA相互作用。YopN的有效分泌依赖于其n端分泌信号（SS）、CBD和SycN/YscB伴侣。在这项研究中，我们研究了YopN CBD在Yop分泌调节中的作用。对YopN SS或SS和CBD被YopE类似区域取代的YopE/YopN杂交蛋白的分析表明，YopN CBD或SycN/YscB伴侣蛋白在YopN分泌中的作用独立于它们在YopN分泌中的作用。为了进一步分析YopN CBD在调控Yop分泌中的作用，在整个YopN CBD中产生了一系列四丙氨酸替代突变体。其中许多突变体在Yop分泌的调节上存在缺陷，但在YopN分泌或YopN与SycN/YscB伴侣的相互作用上没有缺陷。最后，我们建立了条件，使YopN和TyeA能够在缺乏SycN/YscB伴侣的情况下调节Yop的分泌。重要的是，即使在建立的SycN/YscB伴侣独立条件下，许多YopN CBD突变体在调节Yop分泌方面仍保持其缺陷。这些研究表明，YopN的CBD区域在调节Yop分泌中的作用独立于其在YopN分泌或SycN/YscB伴侣结合中的作用。
Numerous Gram-negative bacterial pathogens employ type III secretion systems (T3SSs) to inject effector proteins into eukaryotic cells. The activation of the type III secretion (T3S) process is tightly controlled in all T3SSs. In Yersinia pestis, the secretion of effector proteins, termed Yersinia outer proteins (Yops), is regulated by the activity of the YopN/SycN/YscB/TyeA complex. YopN is a secreted protein that interacts with the SycN/YscB chaperone via an N-terminal chaperone-binding domain (CBD) and with TyeA via a C-terminal TyeA-binding domain (TBD). Efficient YopN secretion is dependent upon its N-terminal secretion signal (SS), CBD, and the SycN/YscB chaperone. In this study, we investigate the role of the YopN CBD in the regulation of Yop secretion. Analysis of YopE/YopN hybrid proteins in which the YopN SS or SS and CBD were replaced with the analogous regions of YopE indicated that the YopN CBD or SycN/YscB chaperone play a role in the regulation of Yop secretion that is independent of their established roles in YopN secretion. To further analyze the role of the YopN CBD in the regulation of Yop secretion a series of tetra-alanine substitution mutants were generated throughout the YopN CBD. A number of these mutants exhibited a defect in the regulation of Yop secretion but showed no defect in YopN secretion or in the interaction of YopN with the SycN/YscB chaperone. Finally, conditions were established that enabled YopN and TyeA to regulate Yop secretion in the absence of the SycN/YscB chaperone. Importantly, a number of the YopN CBD mutants maintained their defect in the regulation of Yop secretion even under the established SycN/YscB chaperone-independent conditions. These studies establish a role for the CBD region of YopN in the regulation of Yop secretion that is independent from its role in YopN secretion or in the binding of the SycN/YscB chaperone.
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影响因子：
3.1
作者：
Bahrani, FK;Sansonetti, PJ;Parsot, C
通讯作者：
Parsot, C
影响因子：
3.6
作者：
Agrain, C;Callebaut, I;Cornelis, GR
通讯作者：
Cornelis, GR
影响因子：
3.1
作者：
Garcia, Julie Torruellas;Ferracci, Franco;Plano, Gregory V.
通讯作者：
Plano, Gregory V.
影响因子：
3.5
作者：
CHEREPANOV, PP;WACKERNAGEL, W
通讯作者：
WACKERNAGEL, W
影响因子：
3.2
作者：
Jackson, MW;Day, JB;Plano, GV
通讯作者：
Plano, GV