Cargando…

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

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 vi...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2887381/
https://www.ncbi.nlm.nih.gov/pubmed/20478027
http://dx.doi.org/10.1186/1742-4690-7-45
_version_ 1782182541729464320
author 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
author_facet 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
author_sort Stirnnagel, Kristin
collection PubMed
description 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.
format Text
id pubmed-2887381
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-28873812010-06-18 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 Retrovirology Research 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. BioMed Central 2010-05-17 /pmc/articles/PMC2887381/ /pubmed/20478027 http://dx.doi.org/10.1186/1742-4690-7-45 Text en Copyright ©2010 Stirnnagel et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
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
title Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles
title_full Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles
title_fullStr Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles
title_full_unstemmed Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles
title_short Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles
title_sort analysis of prototype foamy virus particle-host cell interaction with autofluorescent retroviral particles
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2887381/
https://www.ncbi.nlm.nih.gov/pubmed/20478027
http://dx.doi.org/10.1186/1742-4690-7-45
work_keys_str_mv AT stirnnagelkristin analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT lufteneggerdaniel analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT stangeannett analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT swiersyanka analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT mullerserik analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT rehjuliane analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT stankenicole analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT großearend analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT chiantiasalvatore analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT kellerheiko analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT schwillepetra analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT hanenberghelmut analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT zentgrafhanswalter analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles
AT lindemanndirk analysisofprototypefoamyvirusparticlehostcellinteractionwithautofluorescentretroviralparticles