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Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome
Despite conserved catalytic integration mechanisms, retroviral intasomes composed of integrase (IN) and viral DNA possess diverse structures with variable numbers of IN subunits. To investigate intasome assembly mechanisms, we employed the Rous sarcoma virus (RSV) IN dimer that assembles a precursor...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7955051/ https://www.ncbi.nlm.nih.gov/pubmed/33712691 http://dx.doi.org/10.1038/s42003-021-01855-2 |
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author | Pandey, Krishan K. Bera, Sibes Shi, Ke Rau, Michael J. Oleru, Amarachi V. Fitzpatrick, James A. J. Engelman, Alan N. Aihara, Hideki Grandgenett, Duane P. |
author_facet | Pandey, Krishan K. Bera, Sibes Shi, Ke Rau, Michael J. Oleru, Amarachi V. Fitzpatrick, James A. J. Engelman, Alan N. Aihara, Hideki Grandgenett, Duane P. |
author_sort | Pandey, Krishan K. |
collection | PubMed |
description | Despite conserved catalytic integration mechanisms, retroviral intasomes composed of integrase (IN) and viral DNA possess diverse structures with variable numbers of IN subunits. To investigate intasome assembly mechanisms, we employed the Rous sarcoma virus (RSV) IN dimer that assembles a precursor tetrameric structure in transit to the mature octameric intasome. We determined the structure of RSV octameric intasome stabilized by a HIV-1 IN strand transfer inhibitor using single particle cryo-electron microscopy. The structure revealed significant flexibility of the two non-catalytic distal IN dimers along with previously unrecognized movement of the conserved intasome core, suggesting ordered conformational transitions between intermediates that may be important to capture the target DNA. Single amino acid substitutions within the IN C-terminal domain affected intasome assembly and function in vitro and infectivity of pseudotyped RSV virions. Unexpectedly, 17 C-terminal amino acids of IN were dispensable for virus infection despite regulating the transition of the tetrameric intasome to the octameric form in vitro. We speculate that this region may regulate the binding of highly flexible distal IN dimers to the intasome core to form the octameric complex. Our studies reveal key steps in the assembly of RSV intasomes. |
format | Online Article Text |
id | pubmed-7955051 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-79550512021-03-28 Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome Pandey, Krishan K. Bera, Sibes Shi, Ke Rau, Michael J. Oleru, Amarachi V. Fitzpatrick, James A. J. Engelman, Alan N. Aihara, Hideki Grandgenett, Duane P. Commun Biol Article Despite conserved catalytic integration mechanisms, retroviral intasomes composed of integrase (IN) and viral DNA possess diverse structures with variable numbers of IN subunits. To investigate intasome assembly mechanisms, we employed the Rous sarcoma virus (RSV) IN dimer that assembles a precursor tetrameric structure in transit to the mature octameric intasome. We determined the structure of RSV octameric intasome stabilized by a HIV-1 IN strand transfer inhibitor using single particle cryo-electron microscopy. The structure revealed significant flexibility of the two non-catalytic distal IN dimers along with previously unrecognized movement of the conserved intasome core, suggesting ordered conformational transitions between intermediates that may be important to capture the target DNA. Single amino acid substitutions within the IN C-terminal domain affected intasome assembly and function in vitro and infectivity of pseudotyped RSV virions. Unexpectedly, 17 C-terminal amino acids of IN were dispensable for virus infection despite regulating the transition of the tetrameric intasome to the octameric form in vitro. We speculate that this region may regulate the binding of highly flexible distal IN dimers to the intasome core to form the octameric complex. Our studies reveal key steps in the assembly of RSV intasomes. Nature Publishing Group UK 2021-03-12 /pmc/articles/PMC7955051/ /pubmed/33712691 http://dx.doi.org/10.1038/s42003-021-01855-2 Text en © The Author(s) 2021 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/. |
spellingShingle | Article Pandey, Krishan K. Bera, Sibes Shi, Ke Rau, Michael J. Oleru, Amarachi V. Fitzpatrick, James A. J. Engelman, Alan N. Aihara, Hideki Grandgenett, Duane P. Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome |
title | Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome |
title_full | Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome |
title_fullStr | Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome |
title_full_unstemmed | Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome |
title_short | Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome |
title_sort | cryo-em structure of the rous sarcoma virus octameric cleaved synaptic complex intasome |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7955051/ https://www.ncbi.nlm.nih.gov/pubmed/33712691 http://dx.doi.org/10.1038/s42003-021-01855-2 |
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