title: Hydrogen bonding at a conserved threonine in lentivirus capsid is required for virus replication.
abstract: The N terminus of the capsid protein (CA) undergoes a considerable conformational change when the human immunodeficiency virus (HIV) protease cleaves it free from the Pr55(Gag) polyprotein. This rearrangement is thought to facilitate the establishment of specific CA-CA interactions that are required for the formation of the mature viral core. Substitution of amino acids that are critical for this refolding of the N terminus is generally detrimental to virus replication and mature virion core morphology. Here, we identify a conserved threonine in simian immunodeficiency virus (SIV) CA, T(47)(CA), that is requisite for viral replication. Replacement of T(47)(CA) in the infectious viral clone SIVmac239 with amino acids with different hydrogen-bonding capabilities and analysis of the effects of these substitutions at key steps in the viral life cycle demonstrate that hydrogen bonding at this position is important for virus infectivity and virion release. In the HIV-based homology model of the mature SIV CA N terminus presented in this study, T(47)(CA) forms several hydrogen bonds with a proximal aspartate, D(50)(CA). This model, coupled with strong phenotypic similarities between viral substitution mutants of each of these two residues in all of the virological assays described herein, indicates that hydrogen bonding between T(47)(CA) and D(50)(CA) is likely required for viral replication. As hydrogen bonding between these two residues is present in HIV CA as well, this interaction presents a potential target for antiviral drug design.
authors.affiliation: Department of Comparative Medicine, Johns Hopkins University School of Medicine, 600 N. Wolfe Street, Baltimore, MD 21287, USA.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Rue
authors.forename: Jason W
authors.initials: JW
authors.lastname: Roos
authors.forename: L Mario
authors.initials: LM
authors.lastname: Amzel
authors.forename: Janice E
authors.initials: JE
authors.lastname: Clements
authors.forename: Sheila A
authors.initials: SA
authors.lastname: Barber
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D015497:HIV-1 / Q000737:chemistry; D006801:Humans; D006860:Hydrogen Bonding; D008854:Microscopy, Electron; D008958:Models, Molecular; D016415:Sequence Alignment; D015302:Simian Immunodeficiency Virus / Q000737:chemistry / Q000472:pathogenicity* / Q000502:physiology*; D013912:Threonine / Q000737:chemistry*; D014764:Viral Proteins / Q000378:metabolism; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure; D014779:Virus Replication
chemical_list: D036022:Capsid Proteins; D014764:Viral Proteins; D013912:Threonine