title: Host cofactors and pharmacologic ligands share an essential interface in HIV-1 capsid that is lost upon disassembly.
abstract: The HIV-1 capsid is involved in all infectious steps from reverse transcription to integration site selection, and is the target of multiple host cell and pharmacologic ligands. However, structural studies have been limited to capsid monomers (CA), and the mechanistic basis for how these ligands influence infection is not well understood. Here we show that a multi-subunit interface formed exclusively within CA hexamers mediates binding to linear epitopes within cellular cofactors NUP153 and CPSF6, and is competed for by the antiretroviral compounds PF74 and BI-2. Each ligand is anchored via a shared phenylalanine-glycine (FG) motif to a pocket within the N-terminal domain of one monomer, and all but BI-2 also make essential interactions across the N-terminal domain: C-terminal domain (NTD:CTD) interface to a second monomer. Dissociation of hexamer into CA monomers prevents high affinity interaction with CPSF6 and PF74, and abolishes binding to NUP153. The second interface is conformationally dynamic, but binding of NUP153 or CPSF6 peptides is accommodated by only one conformation. NUP153 and CPSF6 have overlapping binding sites, but each makes unique CA interactions that, when mutated selectively, perturb cofactor dependency. These results reveal that multiple ligands share an overlapping interface in HIV-1 capsid that is lost upon viral disassembly.
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Price
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom.
authors.forename: David A
authors.initials: DA
authors.lastname: Jacques
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom.
authors.forename: William A
authors.initials: WA
authors.lastname: McEwan
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Fletcher
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Essig
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Chin
authors.affiliation: Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Upul D
authors.initials: UD
authors.lastname: Halambage
authors.affiliation: Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Christopher
authors.initials: C
authors.lastname: Aiken
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Division of Protein and Nucleic Acid Chemistry, Cambridge, United Kingdom.
authors.forename: Leo C
authors.initials: LC
authors.lastname: James
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology; D001665:Binding Sites; D002213:Capsid / Q000737:chemistry* / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D015497:HIV-1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008024:Ligands; D008958:Models, Molecular; D008961:Models, Structural; D009154:Mutation; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism*; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000494:pharmacology; D011083:Polycyclic Compounds / Q000494:pharmacology; D058105:Polymerization; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D048348:Reverse Transcription / Q000187:drug effects; D014771:Virion; D039221:mRNA Cleavage and Polyadenylation Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D019380:Anti-HIV Agents; D036022:Capsid Proteins; D007211:Indoles; D008024:Ligands; C079377:NUP153 protein, human; D028861:Nuclear Pore Complex Proteins; C000591221:PF-3450074; D011083:Polycyclic Compounds; C450562:cleavage factor Im, human; C511625:dodecahydro-7-(2-hydroxyethyl)-3a-methyl-1H-benz(e)indene-3-carbonitrile; D039221:mRNA Cleavage and Polyadenylation Factors; D010649:Phenylalanine