Membrane budding by rabies virus matrix- and phosphoprotein

狂犬病病毒基质和磷蛋白的膜出芽

• 批准号: 13166192
• 负责人: Professor Dr. Karl-Klaus Conzelmann
• 金额: --
• 依托单位: Friedrich-Loeffler-Institut - Bundesforschungsinstitut für Tiergesundheit (FLI)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/13166192?language=en
• 关键词: Membrane; budding; rabies; virus; matrix

Assembly and budding of Rabies virus (RV) and related Rhabdoviruses takes place at the plasma membrane. The major driving force of envelopment and membrane budding is the viral matrix protein M which binds membranes, virus cores and glycoproteins, and which is active in membrane bending and pinching off membrane vesicles. The active process of ¿exocytosis¿ involves cellular membrane-associated factors binding to ¿late domains¿ iden-tified in M or M-like proteins of various enveloped viruses. Of the 2 classical late domain motifs identified in RV M, however, none is essential, and RV M budding is independent of the factor VPS4 downstream of ESCRT, in contrast to viruses budding from internal membranes, such as Ebola virus or HIV. This suggests the presence of so far unidentified late domains in RV M, or the use of alternative, so far unknown mechanisms for viral or vesicle exocytosis at the plasma membrane. In the proposed project, we will dissect the multiple functions of RV M by screening a comprehensive bank of M Ala-scan mutants for complementation of defined steps of the life cycle of defective RVs designed by reverse genetics. In a hierarchical system mutants will be identified that are competent in regulating viral RNA synthesis, binding and con-densation of viral cores and glycoproteins, but have discrete defects in (i) membrane bind-ing (ii) ¿sorting, (iii) -bending and (iv) vesicle pinching-off. In comparison with wtM, the mutants will shed light on the identity of M domains and cellular interacting factors and the molecular mechanisms of viral exocytosis at the plasma membrane.

狂犬病病毒(RV)和相关横纹病毒的组装和萌发发生在质膜上。包膜和膜萌发的主要驱动力是病毒基质蛋白M，它结合膜、病毒核心和糖蛋白，并活跃在膜弯曲和夹断膜泡的过程中。胞吐作用的活性过程涉及细胞膜相关因子与不同包膜病毒M蛋白或类M蛋白中识别的晚期结构域结合。然而，在RV M中发现的2个经典的晚期结构域基序中，没有一个是必需的，并且RV M的萌发不依赖于ESCRT下游的Vps4因子，而不是从内膜萌发的病毒，如埃博拉病毒或HIV。这表明RV M中存在迄今未确定的晚期结构域，或使用了另一种迄今尚不清楚的机制来实现病毒或囊泡在质膜上的胞吐。在拟议的项目中，我们将通过筛选M个Ala-Scan突变体来剖析RV M的多种功能，以补充通过反向遗传学设计的有缺陷的RV生命周期的特定步骤。在一个分级系统中，将鉴定出能够调节病毒核心和糖蛋白的RNA合成、结合和凝聚的突变体，但在(I)膜结合(Ii)分离、(Iii)弯曲和(Iv)囊泡夹断方面存在离散缺陷。与WTM相比，这些突变体将有助于阐明M结构域和细胞相互作用因子的同一性以及病毒在质膜上胞吐的分子机制。