Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles

Kristin Stirnnagel, Daniel Lüftenegger, Annett Stange, Anka Swiersy, Erik Müllers, Juliane Reh, Nicole Stanke, Arend Große, Salvatore Chiantia, Heiko Keller, Petra Schwille, Helmut Hanenberg, Hanswalter Zentgraf, Dirk Lindemann

Research output: Contribution to journalArticle

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Abstract

Background: The foamy virus (FV) replication cycle displays several unique features, which set them apart from orthoretroviruses. First, like other B/D type orthoretroviruses, FV capsids preassemble at the centrosome, but more similar to hepadnaviruses, FV budding is strictly dependent on cognate viral glycoprotein coexpression. Second, the unusually broad host range of FV is thought to be due to use of a very common entry receptor present on host cell plasma membranes, because all cell lines tested in vitro so far are permissive.Results: In order to take advantage of modern fluorescent microscopy techniques to study FV replication, we have created FV Gag proteins bearing a variety of protein tags and evaluated these for their ability to support various steps of FV replication. Addition of even small N-terminal HA-tags to FV Gag severely impaired FV particle release. For example, release was completely abrogated by an N-terminal autofluorescent protein (AFP) fusion, despite apparently normal intracellular capsid assembly. In contrast, C-terminal Gag-tags had only minor effects on particle assembly, egress and particle morphogenesis. The infectivity of C-terminal capsid-tagged FV vector particles was reduced up to 100-fold in comparison to wild type; however, infectivity was rescued by coexpression of wild type Gag and assembly of mixed particles. Specific dose-dependent binding of fluorescent FV particles to target cells was demonstrated in an Env-dependent manner, but not binding to target cell-extracted- or synthetic- lipids. Screening of target cells of various origins resulted in the identification of two cell lines, a human erythroid precursor- and a zebrafish- cell line, resistant to FV Env-mediated FV- and HIV-vector transduction.Conclusions: We have established functional, autofluorescent foamy viral particles as a valuable new tool to study FV - host cell interactions using modern fluorescent imaging techniques. Furthermore, we succeeded for the first time in identifying two cell lines resistant to Prototype Foamy Virus Env-mediated gene transfer. Interestingly, both cell lines still displayed FV Env-dependent attachment of fluorescent retroviral particles, implying a post-binding block potentially due to lack of putative FV entry cofactors. These cell lines might ultimately lead to the identification of the currently unknown ubiquitous cellular entry receptor(s) of FVs.

Original languageEnglish (US)
Article number45
JournalRetrovirology
Volume7
DOIs
StatePublished - May 17 2010
Externally publishedYes

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Spumavirus
Cell Communication
Virion
Cell Line
Capsid
Virus Replication
Virus Release
Hepadnaviridae
Cell Membrane
env Genes
gag Gene Products

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Stirnnagel, K., Lüftenegger, D., Stange, A., Swiersy, A., Müllers, E., Reh, J., ... Lindemann, D. (2010). Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles. Retrovirology, 7, [45]. https://doi.org/10.1186/1742-4690-7-45

Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles. / Stirnnagel, Kristin; Lüftenegger, Daniel; Stange, Annett; Swiersy, Anka; Müllers, Erik; Reh, Juliane; Stanke, Nicole; Große, Arend; Chiantia, Salvatore; Keller, Heiko; Schwille, Petra; Hanenberg, Helmut; Zentgraf, Hanswalter; Lindemann, Dirk.

In: Retrovirology, Vol. 7, 45, 17.05.2010.

Research output: Contribution to journalArticle

Stirnnagel, K, Lüftenegger, D, Stange, A, Swiersy, A, Müllers, E, Reh, J, Stanke, N, Große, A, Chiantia, S, Keller, H, Schwille, P, Hanenberg, H, Zentgraf, H & Lindemann, D 2010, 'Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles', Retrovirology, vol. 7, 45. https://doi.org/10.1186/1742-4690-7-45
Stirnnagel, Kristin ; Lüftenegger, Daniel ; Stange, Annett ; Swiersy, Anka ; Müllers, Erik ; Reh, Juliane ; Stanke, Nicole ; Große, Arend ; Chiantia, Salvatore ; Keller, Heiko ; Schwille, Petra ; Hanenberg, Helmut ; Zentgraf, Hanswalter ; Lindemann, Dirk. / Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles. In: Retrovirology. 2010 ; Vol. 7.
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