In vitro, in cellulo and structural characterizations of the interaction between the integrase of Porcine Endogenous Retrovirus A/C and proteins of the BET family.

Autor: Gallay K; INRA, Université Lyon 1, UMR754, Viral Infections Compared Pathology, 69007, Lyon, France; Université de Lyon, 69000, Lyon, France; UMSl3444 Biosciences Gerland Lyon Sud, 69007, Lyon, France., Blot G; ANSES, Ploufragan/Plouzané Laboratory, Viral Genetics and Bio-Security Unit, Université Européenne de Bretagne, Ploufragan, France., Chahpazoff M; Molecular Microbiology and Structural Biochemistry, MMSB-IBCP, UMR 5086 CNRS University of Lyon, 7, passage du Vercors, 69367, Lyon, Cedex 07, France., Yajjou-Hamalian H; Molecular Microbiology and Structural Biochemistry, MMSB-IBCP, UMR 5086 CNRS University of Lyon, 7, passage du Vercors, 69367, Lyon, Cedex 07, France., Confort MP; INRA, Université Lyon 1, UMR754, Viral Infections Compared Pathology, 69007, Lyon, France; Université de Lyon, 69000, Lyon, France; UMSl3444 Biosciences Gerland Lyon Sud, 69007, Lyon, France., De Boisséson C; ANSES, Ploufragan/Plouzané Laboratory, Viral Genetics and Bio-Security Unit, Université Européenne de Bretagne, Ploufragan, France., Leroux A; ANSES, Ploufragan/Plouzané Laboratory, Viral Genetics and Bio-Security Unit, Université Européenne de Bretagne, Ploufragan, France., Luengo C; INRA, Université Lyon 1, UMR754, Viral Infections Compared Pathology, 69007, Lyon, France; Université de Lyon, 69000, Lyon, France; UMSl3444 Biosciences Gerland Lyon Sud, 69007, Lyon, France., Fiorini F; Molecular Microbiology and Structural Biochemistry, MMSB-IBCP, UMR 5086 CNRS University of Lyon, 7, passage du Vercors, 69367, Lyon, Cedex 07, France., Lavigne M; Pasteur Institute, Virology dept., Institut Cochin- Inserm U1016-CNRS UMR8104-Université Paris Descartes, 27, rue du faubourg Saint Jacques, 75014, Paris, France., Chebloune Y; Pathogenesis and Lentiviral Vaccination Laboratory, INRA, Univ Grenoble Alpes, 570 rue de la Chimie, 38400, St Martin d'Hères, France., Gouet P; Molecular Microbiology and Structural Biochemistry, MMSB-IBCP, UMR 5086 CNRS University of Lyon, 7, passage du Vercors, 69367, Lyon, Cedex 07, France., Moreau K; INRA, Université Lyon 1, UMR754, Viral Infections Compared Pathology, 69007, Lyon, France; Université de Lyon, 69000, Lyon, France; UMSl3444 Biosciences Gerland Lyon Sud, 69007, Lyon, France., Blanchard Y; ANSES, Ploufragan/Plouzané Laboratory, Viral Genetics and Bio-Security Unit, Université Européenne de Bretagne, Ploufragan, France. Electronic address: yannick.blanchard@anses.fr., Ronfort C; INRA, Université Lyon 1, UMR754, Viral Infections Compared Pathology, 69007, Lyon, France; Université de Lyon, 69000, Lyon, France; UMSl3444 Biosciences Gerland Lyon Sud, 69007, Lyon, France. Electronic address: corinne.ronfort@inra.fr.
Jazyk: angličtina
Zdroj: Virology [Virology] 2019 Jun; Vol. 532, pp. 69-81. Date of Electronic Publication: 2019 Apr 16.
DOI: 10.1016/j.virol.2019.04.002
Abstrakt: Retroviral integrase (IN) proteins catalyze the permanent integration of the viral genome into host DNA. They can productively recruit cellular proteins, and the human Bromodomain and Extra-Terminal domain (hBET) proteins have been shown to be co-factors for integration of gamma-retroviruses such as Murine Leukemia Virus (MLV) into human cells. By using two-hybrid, co-immunoprecipitation and in vitro interaction assays, we showed that IN of the gamma- Porcine Endogenous Retrovirus-A/C (PERV IN) interacts through its C-terminal domain (CTD) with hBET proteins. We observed that PERV IN interacts with the BRD2, BRD3 and BRD4 proteins in vitro and that the BRD2 protein specifically binds and co-localizes with PERV IN protein in the nucleus of cells. We further mapped the interaction sites to the conserved Extra-Terminal (ET) domain of the hBET proteins and to several amino acids of the of the C-terminal tail of the PERV IN CTD. Finally, we determined the first experimental structure of an IN CTD - BET ET complex from small-angle X-ray scattering data (SAXS). We showed that the two factors assemble as two distinct modules linked by a short loop which confers partial flexibility. The SAXS-restrained model is structurally compatible with the binding of the PERV intasome to BRD2. Altogether, these data confirm the important role of host BET proteins in the gamma-retroviruses' targeting site and efficiency of integration.
(Copyright © 2019. Published by Elsevier Inc.)
Databáze: MEDLINE
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