An N-terminal domain helical motif of Prototype Foamy Virus Gag with dual functions essential for particle egress and viral infectivity

• Published in: Retrovirology 2013-04, Vol.10 (1), p.45-45, Article 45
• Main Authors: Reh, Juliane, Stange, Annett, Götz, Anne, Rönitz, Marlene, Große, Arend, Lindemann, Dirk
• Format: Article
• Language: English
• Subjects: Analysis; Cold; Foamy virus; Gene mutations; Gene Products, env - chemistry; Gene Products, env - metabolism; Gene Products, gag - chemistry; Gene Products, gag - metabolism; Humans; Peptides; Protein Binding; Protein Interaction Domains and Motifs; Protein-protein interactions; Proteins; Spumavirus - physiology; Studies; Virulence; Virus Attachment; Virus Release
• ISSN: 1742-4690; 1742-4690
• DOI: 10.1186/1742-4690-10-45

Foamy viruses (FVs) have developed a unique budding strategy within the retrovirus family. FV release requires co-expression and a highly specific interaction between capsid (Gag) and glycoprotein (Env), which cannot be complemented by heterologous Env proteins. The interaction domain in FV Env has been mapped in greater detail and resides mainly in the N-terminal tip of the cytoplasmic domain of the Env leader peptide subunit. In contrast, the corresponding domain within Gag is less well defined. Previous investigations suggest that it is located within the N-terminal part of the protein. Here we characterized additional Gag interaction determinants of the prototype FV (PFV) isolate using a combination of particle release, GST pull-down and single cycle infectivity analysis assays. Our results demonstrate that a minimal PFV Gag protein comprising the N-terminal 129 aa was released into the supernatant, whereas proteins lacking this domain failed to do so. Fine mapping of domains within the N-terminus of PFV Gag revealed that the N-terminal 10 aa of PFV Gag were dispensable for viral replication. In contrast, larger deletions or structurally deleterious point mutations in C-terminally adjacent sequences predicted to harbor a helical region abolished particle egress and Gag - Env protein interaction. Pull-down assays, using proteins of mammalian and prokaryotic origin, support the previous hypothesis of a direct interaction of both PFV proteins without requirement for cellular cofactors and suggest a potential direct contact of Env through this N-terminal Gag domain. Furthermore, analysis of point mutants within this domain in context of PFV vector particles indicates additional particle release-independent functions for this structure in viral replication by directly affecting virion infectivity. Thus, our results demonstrate not only a critical function of an N-terminal PFV Gag motif for the essential capsid - glycoprotein interaction required for virus budding but also point out additional functions that affect virion infectivity.