title: Proteolytic refolding of the HIV-1 capsid protein amino-terminus facilitates viral core assembly.
abstract: After budding, the human immunodeficiency virus (HIV) must 'mature' into an infectious viral particle. Viral maturation requires proteolytic processing of the Gag polyprotein at the matrix-capsid junction, which liberates the capsid (CA) domain to condense from the spherical protein coat of the immature virus into the conical core of the mature virus. We propose that upon proteolysis, the amino-terminal end of the capsid refolds into a beta-hairpin/helix structure that is stabilized by formation of a salt bridge between the processed amino-terminus (Pro1) and a highly conserved aspartate residue (Asp51). The refolded amino-terminus then creates a new CA-CA interface that is essential for assembling the condensed conical core. Consistent with this model, we found that recombinant capsid proteins with as few as four matrix residues fused to their amino-termini formed spheres in vitro, but that removing these residues refolded the capsid amino-terminus and redirected protein assembly from spheres to cylinders. Moreover, point mutations throughout the putative CA-CA interface blocked capsid assembly in vitro, core assembly in vivo and viral infectivity. Disruption of the conserved amino-terminal capsid salt bridge also abolished the infectivity of Moloney murine leukemia viral particles, suggesting that lenti- and oncoviruses mature via analogous pathways.
authors.affiliation: Department of Biochemistry, University of Utah, Salt Lake City, UT 84132, USA.
authors.forename: U K
authors.initials: UK
authors.lastname: von Schwedler
authors.forename: T L
authors.initials: TL
authors.lastname: Stemmler
authors.forename: V Y
authors.initials: VY
authors.lastname: Klishko
authors.forename: S
authors.initials: S
authors.lastname: Li
authors.forename: K H
authors.initials: KH
authors.lastname: Albertine
authors.forename: D R
authors.initials: DR
authors.lastname: Davis
authors.forename: W I
authors.initials: WI
authors.lastname: Sundquist
mesh_terms: D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry* / Q000235:genetics; D002460:Cell Line; D017124:Conserved Sequence / Q000235:genetics; D015683:Gene Products, gag / Q000737:chemistry; D015497:HIV-1 / Q000737:chemistry / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000502:physiology; D019906:Nuclear Magnetic Resonance, Biomolecular; D017354:Point Mutation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins; D013601:T-Lymphocytes; D014763:Viral Matrix Proteins / Q000737:chemistry; D014771:Virion / Q000648:ultrastructure; D019065:Virus Assembly / Q000502:physiology*
chemical_list: D015683:Gene Products, gag; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins