Eukaryotic SNARE VAMP3 Dynamically Interacts with Multiple Chlamydial Inclusion Membrane Proteins.
Eukaryotic SNARE VAMP3 Dynamically Interacts with Multiple Chlamydial Inclusion Membrane Proteins.
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DOI：
10.1128/iai.00409-20
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发表时间：
2021-01-19
影响因子：
3.1
通讯作者：
Rucks EA
中科院分区：
文献类型：
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作者：
Bui DC;Jorgenson LM;Ouellette SP;Rucks EA
沙眼衣原体是一种专性细胞内病原体，在被称为衣原体包涵体的膜结合空泡内经历双相发育周期。为了促进与宿主细胞的相互作用，衣原体用III型分泌蛋白（称为Incs）修饰包涵体膜。沙眼衣原体是一种专性细胞内病原体，在被称为衣原体包涵体的膜结合空泡内经历双相发育周期。为了促进与宿主细胞的相互作用，衣原体用III型分泌蛋白（称为Incs）修饰包涵体膜。与所有衣原体蛋白一样，Inc是暂时表达的，在发育周期的早期和中期修改衣原体包涵体。VAMP 3和VAMP 4是真核SNARE蛋白，介导膜融合，并被募集到内含物中以促进内含物扩张。他们的招聘需要从头衣原体蛋白质合成在中期发育周期。因此，我们假设VAMP 3和VAMP 4被Inc招募。在衣原体感染的细胞中，识别特异性SNARE蛋白的Inc结合伴侣一直是难以捉摸的。到目前为止，大多数研究衣原体公司和真核生物蛋白质受益于稳定的相互作用的合作伙伴或强大的相互作用，在感染后的特定时间。虽然这些类型的相互作用是已经确定的主要类别，但它们可能是衣原体-宿主相互作用的例外。因此，我们应用两个独立但互补的实验系统来鉴定VAMP的候选衣原体Inc结合伴侣。基于这些结果，我们创建了C.沙眼衣原体血清型L2以诱导表达候选Inc-FLAG蛋白。在衣原体感染的细胞中，我们发现5个Inc与VAMP 3暂时和短暂地相互作用。此外，印加或ct 813表达的缺失改变了VAMP 3在包涵体中的定位。我们的研究第一次证明了某些宿主蛋白质-公司相互作用的短暂性，这些相互作用有助于衣原体的发育周期。
Chlamydia trachomatis, an obligate intracellular pathogen, undergoes a biphasic developmental cycle within a membrane-bound vacuole called the chlamydial inclusion. To facilitate interactions with the host cell, Chlamydia modifies the inclusion membrane with type III secreted proteins, called Incs. Chlamydia trachomatis, an obligate intracellular pathogen, undergoes a biphasic developmental cycle within a membrane-bound vacuole called the chlamydial inclusion. To facilitate interactions with the host cell, Chlamydia modifies the inclusion membrane with type III secreted proteins, called Incs. As with all chlamydial proteins, Incs are temporally expressed, modifying the chlamydial inclusion during the early and mid-developmental cycle. VAMP3 and VAMP4 are eukaryotic SNARE proteins that mediate membrane fusion and are recruited to the inclusion to facilitate inclusion expansion. Their recruitment requires de novo chlamydial protein synthesis during the mid-developmental cycle. Thus, we hypothesize that VAMP3 and VAMP4 are recruited by Incs. In chlamydia-infected cells, identifying Inc binding partners for SNARE proteins specifically has been elusive. To date, most studies examining chlamydial Inc and eukaryotic proteins have benefitted from stable interacting partners or a robust interaction at a specific time postinfection. While these types of interactions are the predominant class that have been identified, they are likely the exception to chlamydia-host interactions. Therefore, we applied two separate but complementary experimental systems to identify candidate chlamydial Inc binding partners for VAMPs. Based on these results, we created transformed strains of C. trachomatis serovar L2 to inducibly express a candidate Inc-FLAG protein. In chlamydia-infected cells, we found that five Incs temporally and transiently interact with VAMP3. Further, loss of incA or ct813 expression altered VAMP3 localization to the inclusion. For the first time, our studies demonstrate the transient nature of certain host protein-Inc interactions that contribute to the chlamydial developmental cycle.