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Multivalent interactions essential for lentiviral integrase function
A multimer of retroviral integrase (IN) synapses viral DNA ends within a stable intasome nucleoprotein complex for integration into a host cell genome. Reconstitution of the intasome from the maedi-visna virus (MVV), an ovine lentivirus, revealed a large assembly containing sixteen IN subunits(1). H...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9065133/ https://www.ncbi.nlm.nih.gov/pubmed/35504909 http://dx.doi.org/10.1038/s41467-022-29928-8 |
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author | Ballandras-Colas, Allison Chivukula, Vidya Gruszka, Dominika T. Shan, Zelin Singh, Parmit K. Pye, Valerie E. McLean, Rebecca K. Bedwell, Gregory J. Li, Wen Nans, Andrea Cook, Nicola J. Fadel, Hind J. Poeschla, Eric M. Griffiths, David J. Vargas, Javier Taylor, Ian A. Lyumkis, Dmitry Yardimci, Hasan Engelman, Alan N. Cherepanov, Peter |
author_facet | Ballandras-Colas, Allison Chivukula, Vidya Gruszka, Dominika T. Shan, Zelin Singh, Parmit K. Pye, Valerie E. McLean, Rebecca K. Bedwell, Gregory J. Li, Wen Nans, Andrea Cook, Nicola J. Fadel, Hind J. Poeschla, Eric M. Griffiths, David J. Vargas, Javier Taylor, Ian A. Lyumkis, Dmitry Yardimci, Hasan Engelman, Alan N. Cherepanov, Peter |
author_sort | Ballandras-Colas, Allison |
collection | PubMed |
description | A multimer of retroviral integrase (IN) synapses viral DNA ends within a stable intasome nucleoprotein complex for integration into a host cell genome. Reconstitution of the intasome from the maedi-visna virus (MVV), an ovine lentivirus, revealed a large assembly containing sixteen IN subunits(1). Herein, we report cryo-EM structures of the lentiviral intasome prior to engagement of target DNA and following strand transfer, refined at 3.4 and 3.5 Å resolution, respectively. The structures elucidate details of the protein-protein and protein-DNA interfaces involved in lentiviral intasome formation. We show that the homomeric interfaces involved in IN hexadecamer formation and the α-helical configuration of the linker connecting the C-terminal and catalytic core domains are critical for MVV IN strand transfer activity in vitro and for virus infectivity. Single-molecule microscopy in conjunction with photobleaching reveals that the MVV intasome can bind a variable number, up to sixteen molecules, of the lentivirus-specific host factor LEDGF/p75. Concordantly, ablation of endogenous LEDGF/p75 results in gross redistribution of MVV integration sites in human and ovine cells. Our data confirm the importance of the expanded architecture observed in cryo-EM studies of lentiviral intasomes and suggest that this organization underlies multivalent interactions with chromatin for integration targeting to active genes. |
format | Online Article Text |
id | pubmed-9065133 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-90651332022-05-04 Multivalent interactions essential for lentiviral integrase function Ballandras-Colas, Allison Chivukula, Vidya Gruszka, Dominika T. Shan, Zelin Singh, Parmit K. Pye, Valerie E. McLean, Rebecca K. Bedwell, Gregory J. Li, Wen Nans, Andrea Cook, Nicola J. Fadel, Hind J. Poeschla, Eric M. Griffiths, David J. Vargas, Javier Taylor, Ian A. Lyumkis, Dmitry Yardimci, Hasan Engelman, Alan N. Cherepanov, Peter Nat Commun Article A multimer of retroviral integrase (IN) synapses viral DNA ends within a stable intasome nucleoprotein complex for integration into a host cell genome. Reconstitution of the intasome from the maedi-visna virus (MVV), an ovine lentivirus, revealed a large assembly containing sixteen IN subunits(1). Herein, we report cryo-EM structures of the lentiviral intasome prior to engagement of target DNA and following strand transfer, refined at 3.4 and 3.5 Å resolution, respectively. The structures elucidate details of the protein-protein and protein-DNA interfaces involved in lentiviral intasome formation. We show that the homomeric interfaces involved in IN hexadecamer formation and the α-helical configuration of the linker connecting the C-terminal and catalytic core domains are critical for MVV IN strand transfer activity in vitro and for virus infectivity. Single-molecule microscopy in conjunction with photobleaching reveals that the MVV intasome can bind a variable number, up to sixteen molecules, of the lentivirus-specific host factor LEDGF/p75. Concordantly, ablation of endogenous LEDGF/p75 results in gross redistribution of MVV integration sites in human and ovine cells. Our data confirm the importance of the expanded architecture observed in cryo-EM studies of lentiviral intasomes and suggest that this organization underlies multivalent interactions with chromatin for integration targeting to active genes. Nature Publishing Group UK 2022-05-03 /pmc/articles/PMC9065133/ /pubmed/35504909 http://dx.doi.org/10.1038/s41467-022-29928-8 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Ballandras-Colas, Allison Chivukula, Vidya Gruszka, Dominika T. Shan, Zelin Singh, Parmit K. Pye, Valerie E. McLean, Rebecca K. Bedwell, Gregory J. Li, Wen Nans, Andrea Cook, Nicola J. Fadel, Hind J. Poeschla, Eric M. Griffiths, David J. Vargas, Javier Taylor, Ian A. Lyumkis, Dmitry Yardimci, Hasan Engelman, Alan N. Cherepanov, Peter Multivalent interactions essential for lentiviral integrase function |
title | Multivalent interactions essential for lentiviral integrase function |
title_full | Multivalent interactions essential for lentiviral integrase function |
title_fullStr | Multivalent interactions essential for lentiviral integrase function |
title_full_unstemmed | Multivalent interactions essential for lentiviral integrase function |
title_short | Multivalent interactions essential for lentiviral integrase function |
title_sort | multivalent interactions essential for lentiviral integrase function |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9065133/ https://www.ncbi.nlm.nih.gov/pubmed/35504909 http://dx.doi.org/10.1038/s41467-022-29928-8 |
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