Role of the pilot protein YscW in the biogenesis of the YscC secretin in Yersinia enterocolitica
Role of the pilot protein YscW in the biogenesis of the YscC secretin in Yersinia enterocolitica
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DOI：
10.1128/jb.186.16.5366-5375.2004
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发表时间：
2004-08-01
影响因子：
3.2
通讯作者：
Koster, M
中科院分区：
文献类型：
--
作者：
Burghout, P;Beckers, F;Koster, M
YscC分泌素是小肠结肠炎耶尔森氏菌III型蛋白分泌系统的主要组分，并在外膜中形成寡聚体结构。在缺乏外膜脂蛋白YscW的突变体中，分泌强烈减少，并且已经提出YscW在分泌素的生物发生中起作用。为了研究促胰液素和这种假定的先导蛋白之间的相互作用，在Y.小肠结肠炎菌株缺乏所有其他成分的分泌机制。YscW表达增加了寡聚YscC的产量，并且是其外膜定位所需的，证实了YscW作为先导蛋白的功能。而分泌素家族的其他成员的先导结合位点已被确定在C末端，一个截短的YscC衍生物缺乏C-末端96个氨基酸残基的功能和稳定的YscW。脉冲追踪实验显示，在YscC寡聚化完成之前需要类似于30分钟。在不存在YscW的情况下，低聚化被延迟并且YscC低聚物的产率被强烈降低。一个unlipidated形式的YscW蛋白是没有功能的，虽然它仍然与分泌素相互作用，并导致错误定位的YscC，即使在野生型YscW的存在下。因此，YscW与未组装的YscC蛋白相互作用，并促进有效的寡聚化，可能在外膜。
The YscC secretin is a major component of the type III protein secretion system of Yersinia enterocolitica and forms an oligomeric structure in the outer membrane. In a mutant lacking the outer membrane lipoprotein YscW, secretion is strongly reduced, and it has been proposed that YscW plays a role in the biogenesis of the secretin. To study the interaction between the secretin and this putative pilot protein, YscC and YscW were produced in trans in a Y. enterocolitica strain lacking all other components of the secretion machinery. YscW expression increased the yield of oligomeric YscC and was required for its outer membrane localization, confirming the function of YscW as a pilot protein. Whereas the pilot-binding site of other members of the secretin family has been identified in the C terminus, a truncated YscC derivative lacking the C-terminal 96 amino acid residues was functional and stabilized by YscW. Pulse-chase experiments revealed that similar to30 min were required before YscC oligomerization was completed. In the absence of YscW, oligomerization was delayed and the yield of YscC oligomers was strongly reduced. An unlipidated form of the YscW protein was not functional, although it still interacted with the secretin and caused mislocalization of YscC even in the presence of wild-type YscW. Hence, YscW interacts with the unassembled YscC protein and facilitates efficient oligomerization, likely at the outer membrane.