title: Biochemical characterization of rous sarcoma virus MA protein interaction with membranes.
abstract: The MA domain of retroviral Gag proteins mediates association with the host cell membrane during assembly. The biochemical nature of this interaction is not well understood. We have used an in vitro flotation assay to directly measure Rous sarcoma virus (RSV) MA-membrane interaction in the absence of host cell factors. The association of purified MA and MA-containing proteins with liposomes of defined composition was electrostatic in nature and depended upon the presence of a biologically relevant concentration of negatively charged lipids. A mutant MA protein known to be unable to promote Gag membrane association and budding in vivo failed to bind to liposomes. These results were supported by computational modeling. The intrinsic affinity of RSV MA for negatively charged membranes appears insufficient to promote efficient plasma membrane binding during assembly. However, an artificially dimerized form of MA bound to liposomes by at least an order of magnitude more tightly than monomeric MA. This result suggests that the clustering of MA domains, via Gag-Gag interactions during virus assembly, drives membrane association in vivo.
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, 360 Biotechnology Building, Ithaca, NY 14853, USA.
authors.forename: Amanda K
authors.initials: AK
authors.lastname: Dalton
authors.forename: Paul S
authors.initials: PS
authors.lastname: Murray
authors.forename: Diana
authors.initials: D
authors.lastname: Murray
authors.forename: Volker M
authors.initials: VM
authors.lastname: Vogt
mesh_terms: D001358:Avian Sarcoma Viruses / Q000378:metabolism / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D017076:Computer-Aided Design; D008081:Liposomes / Q000378:metabolism*; D008958:Models, Molecular; D010750:Phosphoproteins / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D008081:Liposomes; C110263:MA protein, Rous sarcoma virus; D010750:Phosphoproteins; D014763:Viral Matrix Proteins