Stange Annett, Lüftenegger Daniel, Reh Juliane, Weissenhorn Winfried, Lindemann Dirk
Institut für Virologie, Medizinische Fakultät Carl Gustav Carus, Technische Universität Dresden, Fetscherstr. 74, 01307 Dresden, Germany.
J Virol. 2008 Oct;82(20):9858-69. doi: 10.1128/JVI.00949-08. Epub 2008 Aug 6.
Glycoproteins of several viruses have the capacity to induce release of noninfectious, capsidless particulate structures containing only the viral glycoprotein. Such structures are often called subviral particles (SVP). Foamy viruses (FVs), a special type of retroviruses with a replication strategy combining features of both orthoretroviruses and hepadnaviruses, express a glycoprotein (Env) which has the ability to induce SVP release. However, unlike human hepatitis B virus, prototype FV (PFV) naturally secretes only small amounts of SVPs, because ubiquitination of the Env protein seems to suppress the intrinsic capacity for induction of SVP release. In this study, we characterized the structural determinants influencing PFV SVP release, examined the role of specific Env ubiquitination sites in the regulation of this process, and analyzed the requirement of the cellular vacuolar protein sorting (VPS) machinery for SVP egress. We observed that the cytoplasmic and membrane-spanning domains of both the leader peptide (LP) and the transmembrane (TM) subunit harbor essential as well as inhibitory domains. Furthermore, only ubiquitination at the most N-terminal lysine residues (K(14) and K(15)) in LP reduced cell surface expression and suppressed SVP release to wild-type levels. This suggests that interaction of Env with cellular components required for SVP release suppression is effective only when Env is ubiquitinated at these lysine residues but not at others. Finally, SVP release was sensitive to dominant-negative mutants of late components, but not early components, of the cellular VPS machinery. PFV therefore differs from hepatitis B virus in using the same cellular pathway for egress of both virions and SVPs.
几种病毒的糖蛋白能够诱导释放仅含有病毒糖蛋白的无感染性、无衣壳的颗粒结构。此类结构通常被称为亚病毒颗粒(SVP)。泡沫病毒(FV)是一种特殊类型的逆转录病毒,其复制策略兼具正逆转录病毒和嗜肝DNA病毒的特征,表达一种具有诱导SVP释放能力的糖蛋白(Env)。然而,与人类乙型肝炎病毒不同,原型FV(PFV)天然仅分泌少量SVP,因为Env蛋白的泛素化似乎会抑制诱导SVP释放的内在能力。在本研究中,我们对影响PFV SVP释放的结构决定因素进行了表征,研究了特定Env泛素化位点在该过程调控中的作用,并分析了细胞液泡蛋白分选(VPS)机制对SVP释放的需求。我们观察到,前导肽(LP)和跨膜(TM)亚基的细胞质和跨膜结构域均含有必需结构域和抑制结构域。此外,只有LP中最N端赖氨酸残基(K(14)和K(15))处的泛素化会降低细胞表面表达并将SVP释放抑制至野生型水平。这表明,只有当Env在这些赖氨酸残基而非其他残基处发生泛素化时,Env与抑制SVP释放所需的细胞成分之间的相互作用才有效。最后,SVP释放对细胞VPS机制晚期成分而非早期成分的显性负性突变体敏感。因此,PFV在病毒粒子和SVP释放方面利用相同的细胞途径,这与乙型肝炎病毒不同。