Interaction of the Salmonella-containing vacuole with the endocytic recycling system
Interaction of the Salmonella-containing vacuole with the endocytic recycling system
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DOI：
10.1074/jbc.m500358200
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发表时间：
2005-07-01
影响因子：
4.8
通讯作者：
Brumell, JH
中科院分区：
文献类型：
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作者：
Smith, AC;Cirulis, JT;Brumell, JH
在病原体沙门氏菌进入宿主细胞后，大多数细菌驻留在称为含沙门氏菌空泡（SCV）的膜结合区室中。先前的研究已经确定SCV与早期内体短暂相互作用，但仅获得晚期内体/溶酶体蛋白的子集。然而，完整的一套之间的相互作用的SCV和内吞机制尚未得到表征。在这项研究中，我们已经表明，四个特征性的内吞再循环的调节剂存在于SCV入侵后。相互作用的动力学是不同的每个监管机构; ARF 6和Rab 4立即关联，但它们的存在减少感染后60分钟，而syntaxin 13和Rab 11协会在60分钟达到峰值。使用显性负性的方法，我们确定Rab 11调节的循环的CD 44从空泡，但没有影响主要组织相容性复合体（MHC）I类回收。与此相反，syntaxin 13调节MHC I类分子的再循环，但不调节CD 44。我们还确定，成熟的SCV，衡量收购溶酶体相关的膜蛋白-1，放缓时，回收受损。这些研究结果表明，通过内吞再循环系统的蛋白质运动的调节，通过至少两个并行的途径，并与这些途径的有效相互作用是必要的成熟的沙门氏菌的液泡。我们还证明了使用沙门氏菌入侵作为内体回收事件模型的实用性。
Upon entry of the pathogen Salmonella enterica serovar Typhimurium into host cells, the majority of bacteria reside in a membrane-bound compartment called the Salmonella-containing vacuole (SCV). Previous studies have established that the SCV transiently interacts with early endosomes but only acquires a subset of late endosomal/lysosomal proteins. However, the complete set of interactions between the SCV and the endocytic machinery has yet to be characterized. In this study, we have shown that four characterized regulators of endocytic recycling were present on the SCV after invasion. Interaction kinetics were different for each of the regulators; ARF6 and Rab4 associated immediately, but their presence was diminished 60 min post-infection, whereas syntaxin13 and Rab11 association peaked at 60 min. Using a dominant negative approach, we determined that Rab11 regulates the recycling of CD44 from the vacuole but had no effect on major histocompatibility complex (MHC) class I recycling. In contrast, syntaxin13 regulated the recycling of MHC class I but not of CD44. We also determined that maturation of the SCV, measured by the acquisition of lysosomal associated membrane protein-1, slowed when recycling was impaired. These findings suggest that protein movement through the endocytic recycling system is regulated through at least two concurrent pathways and that efficient interaction with these pathways is necessary for maturation of the Salmonella-containing vacuole. We also demonstrate the utility of using Salmonella invasion as a model of endosomal recycling events.