title: Binding of RNA by the alfalfa mosaic virus movement protein is biphasic.
abstract: The movement protein of alfalfa mosaic virus was expressed in Escherichia coli and purified by cation exchange chromatography. The purified protein bound single-stranded RNA cooperatively in a biphasic manner. At protein saturation, RNA/protein complexes (designated 'primary complexes') were detected by a nitrocellulose-retention assay within 1 min of mixing, both at 4 and 22 degrees C. In contrast, an incubation of 30 min at 22 degrees C was necessary to obtain electrophoretically retarded complexes ('stabilized complexes'), containing a large number of protein molecules bound stably to each molecule of RNA. Stabilization did not take place at 4 degrees C. The rate of formation of the primary complexes was strongly dependent on protein concentration, and thus appeared limited by a bimolecular interaction. In contrast, the rate of stabilization was independent of protein concentration, suggesting that this process consisted of a rearrangement of the primary complexes without binding of additional protein molecules. In agreement with this suggestion, the amount of complexed RNA at equilibrium was the same when assayed by nitrocellulose retention and by electrophoretic retardation. The possibility that these peculiar kinetics could be caused by the presence of Tween 20 in the incubation media is discussed.
authors.affiliation: Institut de Biologie Moléculaire des Plantes, Université Louis Pasteur, Strasbourg, France.
authors.forename: F
authors.initials: F
authors.lastname: Schoumacher
authors.forename: M J
authors.initials: MJ
authors.lastname: Gagey
authors.forename: M
authors.initials: M
authors.lastname: Maira
authors.forename: C
authors.initials: C
authors.lastname: Stussi-Garaud
authors.forename: T
authors.initials: T
authors.lastname: Godefroy-Colburn
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli; D000455:Medicago sativa / Q000382:microbiology; D009029:Mosaic Viruses / Q000378:metabolism*; D012313:RNA / Q000138:chemical synthesis / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D012313:RNA