Cryo-EM structure of the Rous sarcoma virus octameric cleaved synaptic complex intasome.

Autor: Pandey KK; Department of Molecular Microbiology and Immunology, School of Medicine, Saint Louis University, St. Louis, MO, USA. krishan.pandey@health.slu.edu., Bera S; Department of Molecular Microbiology and Immunology, School of Medicine, Saint Louis University, St. Louis, MO, USA., Shi K; Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN, USA., Rau MJ; Washington University Center for Cellular Imaging, Washington University School of Medicine, St. Louis, MO, USA., Oleru AV; Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA., Fitzpatrick JAJ; Washington University Center for Cellular Imaging, Washington University School of Medicine, St. Louis, MO, USA.; Departments of Cell Biology & Physiology and Neuroscience, Washington University in St. Louis, School of Medicine, St. Louis, MO, USA.; Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO, USA., Engelman AN; Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA.; Department of Medicine, Harvard Medical School, Boston, MA, USA., Aihara H; Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN, USA. aihar001@umn.edu., Grandgenett DP; Department of Molecular Microbiology and Immunology, School of Medicine, Saint Louis University, St. Louis, MO, USA. Duane.Grandgenett@health.slu.edu.
Jazyk: angličtina
Zdroj: Communications biology [Commun Biol] 2021 Mar 12; Vol. 4 (1), pp. 330. Date of Electronic Publication: 2021 Mar 12.
DOI: 10.1038/s42003-021-01855-2
Abstrakt: 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.
Databáze: MEDLINE
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