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狂犬病病毒是一种囊膜RNA病毒,主要侵害中枢神经系统,引起人和哺乳动物致命性的脑脊髓炎。现有研究表明囊膜病毒颗粒从感染的细胞中出芽释放时会携带许多可能在病毒的复制过程中发挥重要作用的宿主蛋白。尽管先前已报道某些宿主蛋白可掺入到狂犬病病毒颗粒上,但还没有系统地鉴定狂犬病病毒颗粒上的蛋白质组成。为了理解病毒与宿主间的相互作用的分子机制,本研究在病毒培养和蔗糖密度梯度超离心纯化的基础上,采用蛋白质组学方法分析了纯化的狂犬病病毒颗粒(SRV9弱毒疫苗株)上的蛋白质组成。除了检测到狂犬病病毒编码的五个结构蛋白以外,我们还检测到了50个宿主编码的蛋白。按功能可将其分成十类:胞内转运蛋白(14%),分子伴侣(12%),细胞骨架蛋白(24%),信号转导蛋白(8%),转录调节蛋白(12%),钙离子结合蛋白(6%),酶结合蛋白(6%),代谢作用蛋白(2%),泛素化蛋白(2%),其他功能的蛋白(14%)。利用免疫印迹方法对病毒颗粒上的4个宿主蛋白(肌动蛋白,微管蛋白,微丝结合蛋白和热应激同源蛋白70)进行了验证。本研究首次鉴定了狂犬病病毒颗粒上的宿主蛋白组成,有助于进一步研究该病毒复制与感染机制。
Rabies virus is an enveloped RNA virus that attacks the central nervous system and causes fatal encephalomyelitis in humans and mammals. Previous studies have shown that, when released from infected cells, the encapsulated viral particles carry many host proteins that may play an important role in the replication of the virus. Although it has been previously reported that certain host proteins can be incorporated into rabies virus particles, the protein composition on rabies virus particles has not been systematically identified. In order to understand the molecular mechanism of virus-host interaction, we studied the proteome of purified rabies virus particles (SRV9 attenuated vaccine strain) by proteomics based on virus culture and sucrose density gradient ultracentrifugation composition. In addition to detecting five structural proteins encoded by rabies virus, we also detected 50 host-encoded proteins. They are divided into ten categories by function: intracellular transporter (14%), chaperone (12%), cytoskeletal protein (24%), signal transducer (8%), transcriptional regulator (12%), Calcium-binding protein (6%), enzyme-binding protein (6%), metabotropic protein (2%), ubiquitinated protein (2%), and other functional proteins (14%). Four host proteins (actin, tubulin, microfilament binding protein and heat-stress homologous protein 70) on the virion were verified by immunoblotting. This study, for the first time, identified the host protein composition on rabies virus particles, which helps to further investigate the mechanism of replication and infection of the virus.