title: HIV-1 capsid-cyclophilin interactions determine nuclear import pathway, integration targeting and replication efficiency.
abstract: Lentiviruses such as HIV-1 traverse nuclear pore complexes (NPC) and infect terminally differentiated non-dividing cells, but how they do this is unclear. The cytoplasmic NPC protein Nup358/RanBP2 was identified as an HIV-1 co-factor in previous studies. Here we report that HIV-1 capsid (CA) binds directly to the cyclophilin domain of Nup358/RanBP2. Fusion of the Nup358/RanBP2 cyclophilin (Cyp) domain to the tripartite motif of TRIM5 created a novel inhibitor of HIV-1 replication, consistent with an interaction in vivo. In contrast to CypA binding to HIV-1 CA, Nup358 binding is insensitive to inhibition with cyclosporine, allowing contributions from CypA and Nup358 to be distinguished. Inhibition of CypA reduced dependence on Nup358 and the nuclear basket protein Nup153, suggesting that CypA regulates the choice of the nuclear import machinery that is engaged by the virus. HIV-1 cyclophilin-binding mutants CA G89V and P90A favored integration in genomic regions with a higher density of transcription units and associated features than wild type virus. Integration preference of wild type virus in the presence of cyclosporine was similarly altered to regions of higher transcription density. In contrast, HIV-1 CA alterations in another patch on the capsid surface that render the virus less sensitive to Nup358 or TRN-SR2 depletion (CA N74D, N57A) resulted in integration in genomic regions sparse in transcription units. Both groups of CA mutants are impaired in replication in HeLa cells and human monocyte derived macrophages. Our findings link HIV-1 engagement of cyclophilins with both integration targeting and replication efficiency and provide insight into the conservation of viral cyclophilin recruitment.
authors.affiliation: Division of Infection and Immunity, University College London Medical Research Council Centre for Medical Molecular Virology, London, United Kingdom.
authors.forename: Torsten
authors.initials: T
authors.lastname: Schaller
authors.forename: Karen E
authors.initials: KE
authors.lastname: Ocwieja
authors.forename: Jane
authors.initials: J
authors.lastname: Rasaiyaah
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Price
authors.forename: Troy L
authors.initials: TL
authors.lastname: Brady
authors.forename: Shoshannah L
authors.initials: SL
authors.lastname: Roth
authors.forename: Stéphane
authors.initials: S
authors.lastname: Hué
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Fletcher
authors.forename: KyeongEun
authors.initials: K
authors.lastname: Lee
authors.forename: Vineet N
authors.initials: VN
authors.lastname: KewalRamani
authors.forename: Mahdad
authors.initials: M
authors.lastname: Noursadeghi
authors.forename: Richard G
authors.initials: RG
authors.lastname: Jenner
authors.forename: Leo C
authors.initials: LC
authors.lastname: James
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
authors.forename: Greg J
authors.initials: GJ
authors.lastname: Towers
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology; D015153:Blotting, Western; D036022:Capsid Proteins / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology*; D021984:Cyclophilin A / Q000378:metabolism*; D015658:HIV Infections / Q000378:metabolism*; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000821:virology; D018832:Molecular Chaperones / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014779:Virus Replication / Q000502:physiology
chemical_list: D036022:Capsid Proteins; D018832:Molecular Chaperones; D028861:Nuclear Pore Complex Proteins; C094493:ran-binding protein 2; D021984:Cyclophilin A