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Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection
HIV-1 exploits numerous host cellular pathways for productive infection. To identify novel factors involved in HIV-1 replication, HIV-1 integrase and matrix protein complexes were captured at 4 hours post infection for proteomic analysis using an affinity purification system. Leucine-rich PPR-motif...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3395635/ https://www.ncbi.nlm.nih.gov/pubmed/22808186 http://dx.doi.org/10.1371/journal.pone.0040537 |
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author | Schweitzer, Cameron J. Matthews, John M. Madson, Christian J. Donnellan, Meghan R. Cerny, Ronald L. Belshan, Michael |
author_facet | Schweitzer, Cameron J. Matthews, John M. Madson, Christian J. Donnellan, Meghan R. Cerny, Ronald L. Belshan, Michael |
author_sort | Schweitzer, Cameron J. |
collection | PubMed |
description | HIV-1 exploits numerous host cellular pathways for productive infection. To identify novel factors involved in HIV-1 replication, HIV-1 integrase and matrix protein complexes were captured at 4 hours post infection for proteomic analysis using an affinity purification system. Leucine-rich PPR-motif containing (LRPPRC) protein, a cellular protein involved in mitochondrial function, cell metabolism, and cell-cycle progression was identified as one of the candidate HIV-1 factors. Co-immunoprecipitation RT-PCR experiments confirmed that LRPPRC associated with HIV-1 nucleic acids during the early steps of virus infection. To establish if LRPPRC was critical for HIV-1 infection, three independent LRPPRC knockdown cell lines were constructed (2.7, 3.6, and 4.1). Subcellular fractionation of these cell lines revealed differential knockdown of LRPPRC in subcellular compartments. LRPPRC was knocked down in the insoluble/cytoskeletal fractions of all three cell lines, but the 3.6 and 4.1 cells also showed a reduction in nuclear LRPPRC. Additionally, several cellular factors were downregulated and/or disrupted by loss of LRPPRC. HIV-1 infection was reduced in all three cell lines, but virus production and RNA encapsidation were unaffected, suggesting that LRPPRC was critical for the afferent stage of virus replication. Two of the three cell lines (3.6, 4.1) were refractory for murine leukemia virus infection, a virus dependent on cellular proliferation for productive infection. Consistent with this, these two cell lines exhibited reduced cellular growth with no loss of cellular viability or change in cell cycle phenotype. The early steps of virus infection were also differentially affected among the cell lines. A reduced level of preintegration complex formation was observed in all three cell lines, but viral DNA nuclear import was reduced only in the 3.6 and 4.1 cells. Combined, these data identify LRPPRC as a HIV-1 factor that is involved in HIV-1 replication through more than one mechanism. |
format | Online Article Text |
id | pubmed-3395635 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33956352012-07-17 Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection Schweitzer, Cameron J. Matthews, John M. Madson, Christian J. Donnellan, Meghan R. Cerny, Ronald L. Belshan, Michael PLoS One Research Article HIV-1 exploits numerous host cellular pathways for productive infection. To identify novel factors involved in HIV-1 replication, HIV-1 integrase and matrix protein complexes were captured at 4 hours post infection for proteomic analysis using an affinity purification system. Leucine-rich PPR-motif containing (LRPPRC) protein, a cellular protein involved in mitochondrial function, cell metabolism, and cell-cycle progression was identified as one of the candidate HIV-1 factors. Co-immunoprecipitation RT-PCR experiments confirmed that LRPPRC associated with HIV-1 nucleic acids during the early steps of virus infection. To establish if LRPPRC was critical for HIV-1 infection, three independent LRPPRC knockdown cell lines were constructed (2.7, 3.6, and 4.1). Subcellular fractionation of these cell lines revealed differential knockdown of LRPPRC in subcellular compartments. LRPPRC was knocked down in the insoluble/cytoskeletal fractions of all three cell lines, but the 3.6 and 4.1 cells also showed a reduction in nuclear LRPPRC. Additionally, several cellular factors were downregulated and/or disrupted by loss of LRPPRC. HIV-1 infection was reduced in all three cell lines, but virus production and RNA encapsidation were unaffected, suggesting that LRPPRC was critical for the afferent stage of virus replication. Two of the three cell lines (3.6, 4.1) were refractory for murine leukemia virus infection, a virus dependent on cellular proliferation for productive infection. Consistent with this, these two cell lines exhibited reduced cellular growth with no loss of cellular viability or change in cell cycle phenotype. The early steps of virus infection were also differentially affected among the cell lines. A reduced level of preintegration complex formation was observed in all three cell lines, but viral DNA nuclear import was reduced only in the 3.6 and 4.1 cells. Combined, these data identify LRPPRC as a HIV-1 factor that is involved in HIV-1 replication through more than one mechanism. Public Library of Science 2012-07-12 /pmc/articles/PMC3395635/ /pubmed/22808186 http://dx.doi.org/10.1371/journal.pone.0040537 Text en Schweitzer et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Schweitzer, Cameron J. Matthews, John M. Madson, Christian J. Donnellan, Meghan R. Cerny, Ronald L. Belshan, Michael Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection |
title | Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection |
title_full | Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection |
title_fullStr | Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection |
title_full_unstemmed | Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection |
title_short | Knockdown of the Cellular Protein LRPPRC Attenuates HIV-1 Infection |
title_sort | knockdown of the cellular protein lrpprc attenuates hiv-1 infection |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3395635/ https://www.ncbi.nlm.nih.gov/pubmed/22808186 http://dx.doi.org/10.1371/journal.pone.0040537 |
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