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Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection

The role of accessory proteins during cell-to-cell transmission of HIV-1 has not been explicitly defined. In part, this is related to difficulties in measuring virus replication in cell cocultures with high accuracy, as cells coexist at different stages of infection and separation of effector cells...

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Autores principales: Zotova, Anastasia, Atemasova, Anastasia, Pichugin, Alexey, Filatov, Alexander, Mazurov, Dmitriy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6563509/
https://www.ncbi.nlm.nih.gov/pubmed/31027334
http://dx.doi.org/10.3390/v11050390
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author Zotova, Anastasia
Atemasova, Anastasia
Pichugin, Alexey
Filatov, Alexander
Mazurov, Dmitriy
author_facet Zotova, Anastasia
Atemasova, Anastasia
Pichugin, Alexey
Filatov, Alexander
Mazurov, Dmitriy
author_sort Zotova, Anastasia
collection PubMed
description The role of accessory proteins during cell-to-cell transmission of HIV-1 has not been explicitly defined. In part, this is related to difficulties in measuring virus replication in cell cocultures with high accuracy, as cells coexist at different stages of infection and separation of effector cells from target cells is complicated. In this study, we used replication-dependent reporter vectors to determine requirements for Vif, Vpu, Vpr, or Nef during one cycle of HIV-1 cell coculture and cell-free infection in lymphoid and nonlymphoid cells. Comparative analysis of HIV-1 replication in two cell systems showed that, irrespective of transmission way, accessory proteins were generally less required for virus replication in 293T/CD4/X4 cells than in Jurkat-to-Raji/CD4 cell cocultures. This is consistent with a well-established fact that lymphoid cells express a broad spectrum of restriction factors, while nonlymphoid cells are rather limited in this regard. Remarkably, Vpu deletion reduced the level of cell-free infection, but enhanced the level of cell coculture infection and increased the fraction of multiply infected cells. Nef deficiency did not influence or moderately reduced HIV-1 infection in nonlymphoid and lymphoid cell cocultures, respectively, but strongly affected cell-free infection. Knockout of BST2—a Vpu antagonizing restriction factor—in Jurkat producer cells abolished the enhanced replication of HIV-1 ΔVpu in cell coculture and prevented the formation of viral clusters on cell surface. Thus, BST2-tethered viral particles mediated cell coculture infection more efficiently and at a higher level of multiplicity than diffusely distributed virions. In conclusion, our results demonstrate that the mode of transmission may determine the degree of accessory protein requirements during HIV-1 infection.
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spelling pubmed-65635092019-06-17 Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection Zotova, Anastasia Atemasova, Anastasia Pichugin, Alexey Filatov, Alexander Mazurov, Dmitriy Viruses Article The role of accessory proteins during cell-to-cell transmission of HIV-1 has not been explicitly defined. In part, this is related to difficulties in measuring virus replication in cell cocultures with high accuracy, as cells coexist at different stages of infection and separation of effector cells from target cells is complicated. In this study, we used replication-dependent reporter vectors to determine requirements for Vif, Vpu, Vpr, or Nef during one cycle of HIV-1 cell coculture and cell-free infection in lymphoid and nonlymphoid cells. Comparative analysis of HIV-1 replication in two cell systems showed that, irrespective of transmission way, accessory proteins were generally less required for virus replication in 293T/CD4/X4 cells than in Jurkat-to-Raji/CD4 cell cocultures. This is consistent with a well-established fact that lymphoid cells express a broad spectrum of restriction factors, while nonlymphoid cells are rather limited in this regard. Remarkably, Vpu deletion reduced the level of cell-free infection, but enhanced the level of cell coculture infection and increased the fraction of multiply infected cells. Nef deficiency did not influence or moderately reduced HIV-1 infection in nonlymphoid and lymphoid cell cocultures, respectively, but strongly affected cell-free infection. Knockout of BST2—a Vpu antagonizing restriction factor—in Jurkat producer cells abolished the enhanced replication of HIV-1 ΔVpu in cell coculture and prevented the formation of viral clusters on cell surface. Thus, BST2-tethered viral particles mediated cell coculture infection more efficiently and at a higher level of multiplicity than diffusely distributed virions. In conclusion, our results demonstrate that the mode of transmission may determine the degree of accessory protein requirements during HIV-1 infection. MDPI 2019-04-26 /pmc/articles/PMC6563509/ /pubmed/31027334 http://dx.doi.org/10.3390/v11050390 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zotova, Anastasia
Atemasova, Anastasia
Pichugin, Alexey
Filatov, Alexander
Mazurov, Dmitriy
Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection
title Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection
title_full Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection
title_fullStr Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection
title_full_unstemmed Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection
title_short Distinct Requirements for HIV-1 Accessory Proteins during Cell Coculture and Cell-Free Infection
title_sort distinct requirements for hiv-1 accessory proteins during cell coculture and cell-free infection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6563509/
https://www.ncbi.nlm.nih.gov/pubmed/31027334
http://dx.doi.org/10.3390/v11050390
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