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The autophagy protein ATG9A promotes HIV-1 infectivity
BACKGROUND: Nef is a multifunctional accessory protein encoded by HIV-1, HIV-2 and SIV that plays critical roles in viral pathogenesis, contributing to viral replication, assembly, budding, infectivity and immune evasion, through engagement of various host cell pathways. RESULTS: To gain a better un...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6607583/ https://www.ncbi.nlm.nih.gov/pubmed/31269971 http://dx.doi.org/10.1186/s12977-019-0480-3 |
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author | Mailler, Elodie Waheed, Abdul A. Park, Sang-Yoon Gershlick, David C. Freed, Eric O. Bonifacino, Juan S. |
author_facet | Mailler, Elodie Waheed, Abdul A. Park, Sang-Yoon Gershlick, David C. Freed, Eric O. Bonifacino, Juan S. |
author_sort | Mailler, Elodie |
collection | PubMed |
description | BACKGROUND: Nef is a multifunctional accessory protein encoded by HIV-1, HIV-2 and SIV that plays critical roles in viral pathogenesis, contributing to viral replication, assembly, budding, infectivity and immune evasion, through engagement of various host cell pathways. RESULTS: To gain a better understanding of the role of host proteins in the functions of Nef, we carried out tandem affinity purification-mass spectrometry analysis, and identified over 70 HIV-1 Nef-interacting proteins, including the autophagy-related 9A (ATG9A) protein. ATG9A is a transmembrane component of the machinery for autophagy, a catabolic process in which cytoplasmic components are degraded in lysosomal compartments. Pulldown experiments demonstrated that ATG9A interacts with Nef from not only HIV-1 and but also SIV (cpz, smm and mac). However, expression of HIV-1 Nef had no effect on the levels and localization of ATG9A, and on autophagy, in the host cells. To investigate a possible role for ATG9A in virus replication, we knocked out ATG9A in HeLa cervical carcinoma and Jurkat T cells, and analyzed virus release and infectivity. We observed that ATG9A knockout (KO) had no effect on the release of wild-type (WT) or Nef-defective HIV-1 in these cells. However, the infectivity of WT virus produced from ATG9A-KO HeLa and Jurkat cells was reduced by ~ fourfold and eightfold, respectively, relative to virus produced from WT cells. This reduction in infectivity was independent of the interaction of Nef with ATG9A, and was not due to reduced incorporation of the viral envelope (Env) glycoprotein into the virus. The loss of HIV-1 infectivity was rescued by pseudotyping HIV-1 virions with the vesicular stomatitis virus G glycoprotein. CONCLUSIONS: These studies indicate that ATG9A promotes HIV-1 infectivity in an Env-dependent manner. The interaction of Nef with ATG9A, however, is not required for Nef to enhance HIV-1 infectivity. We speculate that ATG9A could promote infectivity by participating in either the removal of a factor that inhibits infectivity or the incorporation of a factor that enhances infectivity of the viral particles. These studies thus identify a novel host cell factor implicated in HIV-1 infectivity, which may be amenable to pharmacologic manipulation for treatment of HIV-1 infection. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12977-019-0480-3) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6607583 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-66075832019-07-12 The autophagy protein ATG9A promotes HIV-1 infectivity Mailler, Elodie Waheed, Abdul A. Park, Sang-Yoon Gershlick, David C. Freed, Eric O. Bonifacino, Juan S. Retrovirology Research BACKGROUND: Nef is a multifunctional accessory protein encoded by HIV-1, HIV-2 and SIV that plays critical roles in viral pathogenesis, contributing to viral replication, assembly, budding, infectivity and immune evasion, through engagement of various host cell pathways. RESULTS: To gain a better understanding of the role of host proteins in the functions of Nef, we carried out tandem affinity purification-mass spectrometry analysis, and identified over 70 HIV-1 Nef-interacting proteins, including the autophagy-related 9A (ATG9A) protein. ATG9A is a transmembrane component of the machinery for autophagy, a catabolic process in which cytoplasmic components are degraded in lysosomal compartments. Pulldown experiments demonstrated that ATG9A interacts with Nef from not only HIV-1 and but also SIV (cpz, smm and mac). However, expression of HIV-1 Nef had no effect on the levels and localization of ATG9A, and on autophagy, in the host cells. To investigate a possible role for ATG9A in virus replication, we knocked out ATG9A in HeLa cervical carcinoma and Jurkat T cells, and analyzed virus release and infectivity. We observed that ATG9A knockout (KO) had no effect on the release of wild-type (WT) or Nef-defective HIV-1 in these cells. However, the infectivity of WT virus produced from ATG9A-KO HeLa and Jurkat cells was reduced by ~ fourfold and eightfold, respectively, relative to virus produced from WT cells. This reduction in infectivity was independent of the interaction of Nef with ATG9A, and was not due to reduced incorporation of the viral envelope (Env) glycoprotein into the virus. The loss of HIV-1 infectivity was rescued by pseudotyping HIV-1 virions with the vesicular stomatitis virus G glycoprotein. CONCLUSIONS: These studies indicate that ATG9A promotes HIV-1 infectivity in an Env-dependent manner. The interaction of Nef with ATG9A, however, is not required for Nef to enhance HIV-1 infectivity. We speculate that ATG9A could promote infectivity by participating in either the removal of a factor that inhibits infectivity or the incorporation of a factor that enhances infectivity of the viral particles. These studies thus identify a novel host cell factor implicated in HIV-1 infectivity, which may be amenable to pharmacologic manipulation for treatment of HIV-1 infection. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12977-019-0480-3) contains supplementary material, which is available to authorized users. BioMed Central 2019-07-03 /pmc/articles/PMC6607583/ /pubmed/31269971 http://dx.doi.org/10.1186/s12977-019-0480-3 Text en © The Author(s) 2019 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 Mailler, Elodie Waheed, Abdul A. Park, Sang-Yoon Gershlick, David C. Freed, Eric O. Bonifacino, Juan S. The autophagy protein ATG9A promotes HIV-1 infectivity |
title | The autophagy protein ATG9A promotes HIV-1 infectivity |
title_full | The autophagy protein ATG9A promotes HIV-1 infectivity |
title_fullStr | The autophagy protein ATG9A promotes HIV-1 infectivity |
title_full_unstemmed | The autophagy protein ATG9A promotes HIV-1 infectivity |
title_short | The autophagy protein ATG9A promotes HIV-1 infectivity |
title_sort | autophagy protein atg9a promotes hiv-1 infectivity |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6607583/ https://www.ncbi.nlm.nih.gov/pubmed/31269971 http://dx.doi.org/10.1186/s12977-019-0480-3 |
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