title: Kinetic analysis of the role of intersubunit interactions in human immunodeficiency virus type 1 capsid protein assembly in vitro.
abstract: The human immunodeficiency virus type 1 (HIV-1) capsid protein (CA) plays a crucial role in both assembly and maturation of the virion. Numerous recent studies have focused on either the soluble form of CA or the polymer end product of in vitro CA assembly. The CA polymer, in particular, has been used to study CA-CA interactions because it is a good model for the CA interactions within the virion core. However, analysis of the process of in vitro CA assembly can yield valuable insights into CA-CA interactions and the mechanism of core assembly. We describe here a method for the analysis of CA assembly kinetics wherein the progress of assembly is monitored by using turbidity. At pH 7.0 the addition of either of the isolated CA domains (i.e., the N or the C domain) to an assembly reaction caused a decrease in the assembly rate by competing for binding to the full-length CA protein. At pH 8.0 the addition of the isolated C domain had a similar inhibitory affect on CA assembly. However, at pH 8.0 the isolated N domain had no affect on the rate of CA assembly but, when mixed with the C domain, it alleviated the C-domain inhibition. These data provide biochemical evidence for a pH-sensitive homotypic N-domain interaction, as well as for an N- and C-domain interaction.
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294-2170, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Lanman
authors.forename: Jennifer
authors.initials: J
authors.lastname: Sexton
authors.forename: Michael
authors.initials: M
authors.lastname: Sakalian
authors.forename: Peter E
authors.initials: PE
authors.lastname: Prevelige
mesh_terms: D002213:Capsid / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D015497:HIV-1 / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008854:Microscopy, Electron; D012965:Sodium Chloride; D019065:Virus Assembly
chemical_list: D012965:Sodium Chloride