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Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function
BACKGROUND: HIV-1 Vif interacts with the cellular core-binding factor β (CBFβ) and counteracts the protective roles of certain human APOBEC3 (A3) proteins by targeting them for proteasomal degradation. Previous studies have identified some amino acids important for Vif–CBFβ interactions, and recentl...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5356271/ https://www.ncbi.nlm.nih.gov/pubmed/28302150 http://dx.doi.org/10.1186/s12977-017-0346-5 |
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author | Desimmie, Belete A. Smith, Jessica L. Matsuo, Hiroshi Hu, Wei-Shau Pathak, Vinay K. |
author_facet | Desimmie, Belete A. Smith, Jessica L. Matsuo, Hiroshi Hu, Wei-Shau Pathak, Vinay K. |
author_sort | Desimmie, Belete A. |
collection | PubMed |
description | BACKGROUND: HIV-1 Vif interacts with the cellular core-binding factor β (CBFβ) and counteracts the protective roles of certain human APOBEC3 (A3) proteins by targeting them for proteasomal degradation. Previous studies have identified some amino acids important for Vif–CBFβ interactions, and recently a co-crystal structure of a pentameric complex of HIV-1 Vif, CBFβ, Cul5, EloB, and EloC was resolved. However, a comprehensive analysis of Vif–CBFβ interactions that are important for Vif function has not been performed. RESULTS: Here, we carried out double-alanine scanning mutagenesis of the first 60 amino acids of Vif and determined their effects on interaction with CBFβ and their ability to induce A3G degradation as well as rescue HIV-1 replication in the presence of A3G. We found that multiple Vif residues are involved in the extensive N-terminal Vif–CBFβ interaction and that the (5)WQVMIVW(11) region of Vif is the major determinant. A minimum of three alanine substitutions are required to completely abrogate the Vif–CBFβ interaction and Vif’s ability to rescue HIV-1 infectivity in the presence of A3G. Mutational analysis of CBFβ revealed that F68 and I55 residues are important and participate in a tripartite hydrophobic interaction with W5 of Vif to maintain a stable and functional Vif–CBFβ complex. We also determined that CBFβ amino acids (73)WQGEQR(78), which are not resolved in the structure of the pentameric complex, are not involved in interaction with HIV-1 Vif. CONCLUSIONS: Our results provide detailed insight into the Vif–CBFβ interactions that are critical for Vif function and may contribute to the rational design of HIV-1 inhibitors that block Vif-mediated degradation of A3 proteins. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12977-017-0346-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5356271 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-53562712017-03-22 Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function Desimmie, Belete A. Smith, Jessica L. Matsuo, Hiroshi Hu, Wei-Shau Pathak, Vinay K. Retrovirology Research BACKGROUND: HIV-1 Vif interacts with the cellular core-binding factor β (CBFβ) and counteracts the protective roles of certain human APOBEC3 (A3) proteins by targeting them for proteasomal degradation. Previous studies have identified some amino acids important for Vif–CBFβ interactions, and recently a co-crystal structure of a pentameric complex of HIV-1 Vif, CBFβ, Cul5, EloB, and EloC was resolved. However, a comprehensive analysis of Vif–CBFβ interactions that are important for Vif function has not been performed. RESULTS: Here, we carried out double-alanine scanning mutagenesis of the first 60 amino acids of Vif and determined their effects on interaction with CBFβ and their ability to induce A3G degradation as well as rescue HIV-1 replication in the presence of A3G. We found that multiple Vif residues are involved in the extensive N-terminal Vif–CBFβ interaction and that the (5)WQVMIVW(11) region of Vif is the major determinant. A minimum of three alanine substitutions are required to completely abrogate the Vif–CBFβ interaction and Vif’s ability to rescue HIV-1 infectivity in the presence of A3G. Mutational analysis of CBFβ revealed that F68 and I55 residues are important and participate in a tripartite hydrophobic interaction with W5 of Vif to maintain a stable and functional Vif–CBFβ complex. We also determined that CBFβ amino acids (73)WQGEQR(78), which are not resolved in the structure of the pentameric complex, are not involved in interaction with HIV-1 Vif. CONCLUSIONS: Our results provide detailed insight into the Vif–CBFβ interactions that are critical for Vif function and may contribute to the rational design of HIV-1 inhibitors that block Vif-mediated degradation of A3 proteins. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12977-017-0346-5) contains supplementary material, which is available to authorized users. BioMed Central 2017-03-17 /pmc/articles/PMC5356271/ /pubmed/28302150 http://dx.doi.org/10.1186/s12977-017-0346-5 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Desimmie, Belete A. Smith, Jessica L. Matsuo, Hiroshi Hu, Wei-Shau Pathak, Vinay K. Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function |
title | Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function |
title_full | Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function |
title_fullStr | Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function |
title_full_unstemmed | Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function |
title_short | Identification of a tripartite interaction between the N-terminus of HIV-1 Vif and CBFβ that is critical for Vif function |
title_sort | identification of a tripartite interaction between the n-terminus of hiv-1 vif and cbfβ that is critical for vif function |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5356271/ https://www.ncbi.nlm.nih.gov/pubmed/28302150 http://dx.doi.org/10.1186/s12977-017-0346-5 |
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