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Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex
HIV-1 Rev mediates the nuclear export of intron-containing viral RNA transcripts and is essential for viral replication. Rev is imported into the nucleus by the host protein importin β (Impβ), but how Rev associates with Impβ is poorly understood. Here, we report biochemical, mutational, and biophys...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Life Science Alliance LLC
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9396022/ https://www.ncbi.nlm.nih.gov/pubmed/35995566 http://dx.doi.org/10.26508/lsa.202201431 |
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author | Spittler, Didier Indorato, Rose-Laure Boeri Erba, Elisabetta Delaforge, Elise Signor, Luca Harris, Simon J Garcia-Saez, Isabel Palencia, Andrés Gabel, Frank Blackledge, Martin Noirclerc-Savoye, Marjolaine Petosa, Carlo |
author_facet | Spittler, Didier Indorato, Rose-Laure Boeri Erba, Elisabetta Delaforge, Elise Signor, Luca Harris, Simon J Garcia-Saez, Isabel Palencia, Andrés Gabel, Frank Blackledge, Martin Noirclerc-Savoye, Marjolaine Petosa, Carlo |
author_sort | Spittler, Didier |
collection | PubMed |
description | HIV-1 Rev mediates the nuclear export of intron-containing viral RNA transcripts and is essential for viral replication. Rev is imported into the nucleus by the host protein importin β (Impβ), but how Rev associates with Impβ is poorly understood. Here, we report biochemical, mutational, and biophysical studies of the Impβ/Rev complex. We show that Impβ binds two Rev monomers through independent binding sites, in contrast to the 1:1 binding stoichiometry observed for most Impβ cargos. Peptide scanning data and charge-reversal mutations identify the N-terminal tip of Rev helix α2 within Rev’s arginine-rich motif (ARM) as a primary Impβ-binding epitope. Cross-linking mass spectrometry and compensatory mutagenesis data combined with molecular docking simulations suggest a structural model in which one Rev monomer binds to the C-terminal half of Impβ with Rev helix α2 roughly parallel to the HEAT-repeat superhelical axis, whereas the other monomer binds to the N-terminal half. These findings shed light on the molecular basis of Rev recognition by Impβ and highlight an atypical binding behavior that distinguishes Rev from canonical cellular Impβ cargos. |
format | Online Article Text |
id | pubmed-9396022 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Life Science Alliance LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-93960222022-09-14 Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex Spittler, Didier Indorato, Rose-Laure Boeri Erba, Elisabetta Delaforge, Elise Signor, Luca Harris, Simon J Garcia-Saez, Isabel Palencia, Andrés Gabel, Frank Blackledge, Martin Noirclerc-Savoye, Marjolaine Petosa, Carlo Life Sci Alliance Research Articles HIV-1 Rev mediates the nuclear export of intron-containing viral RNA transcripts and is essential for viral replication. Rev is imported into the nucleus by the host protein importin β (Impβ), but how Rev associates with Impβ is poorly understood. Here, we report biochemical, mutational, and biophysical studies of the Impβ/Rev complex. We show that Impβ binds two Rev monomers through independent binding sites, in contrast to the 1:1 binding stoichiometry observed for most Impβ cargos. Peptide scanning data and charge-reversal mutations identify the N-terminal tip of Rev helix α2 within Rev’s arginine-rich motif (ARM) as a primary Impβ-binding epitope. Cross-linking mass spectrometry and compensatory mutagenesis data combined with molecular docking simulations suggest a structural model in which one Rev monomer binds to the C-terminal half of Impβ with Rev helix α2 roughly parallel to the HEAT-repeat superhelical axis, whereas the other monomer binds to the N-terminal half. These findings shed light on the molecular basis of Rev recognition by Impβ and highlight an atypical binding behavior that distinguishes Rev from canonical cellular Impβ cargos. Life Science Alliance LLC 2022-08-22 /pmc/articles/PMC9396022/ /pubmed/35995566 http://dx.doi.org/10.26508/lsa.202201431 Text en © 2022 Spittler et al. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Articles Spittler, Didier Indorato, Rose-Laure Boeri Erba, Elisabetta Delaforge, Elise Signor, Luca Harris, Simon J Garcia-Saez, Isabel Palencia, Andrés Gabel, Frank Blackledge, Martin Noirclerc-Savoye, Marjolaine Petosa, Carlo Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex |
title | Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex |
title_full | Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex |
title_fullStr | Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex |
title_full_unstemmed | Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex |
title_short | Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex |
title_sort | binding stoichiometry and structural model of the hiv-1 rev/importin β complex |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9396022/ https://www.ncbi.nlm.nih.gov/pubmed/35995566 http://dx.doi.org/10.26508/lsa.202201431 |
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