title: Unphosphorylated rhabdoviridae phosphoproteins form elongated dimers in solution.
abstract: The phosphoprotein (P) is an essential component of the replication machinery of rabies virus (RV) and vesicular stomatitis virus (VSV), and the oligomerization of P, potentially controlled by phosphorylation, is required for its function. Up to now the stoichiometry of phosphoprotein oligomers has been controversial. Size exclusion chromatography combined with detection by multiangle laser light scattering shows that the recombinant unphosphorylated phosphoproteins from VSV and from RV exist as dimers in solution. Hydrodynamic analysis indicates that the dimers are highly asymmetric, with a Stokes radius of 4.8-5.3 nm and a frictional ratio larger than 1.7. Small-angle neutron scattering experiments confirm the dimeric state and the asymmetry of the structure and yield a radius of gyration of about 5.3 nm and a cross-sectional radius of gyration of about 1.6-1.8 nm. Similar hydrodynamic properties and molecular dimensions were obtained with a variant of VSV phosphoprotein in which Ser60 and Thr62 are substituted by Asp residues and which has been reported previously to mimic phosphorylation by inducing oligomerization and activating transcription. Here, we show that this mutant also forms a dimer with hydrodynamic properties and molecular dimensions similar to those of the wild type protein. However, incubation at 30 degrees C for several hours induced self-assembly of both wild type and mutant proteins, leading to the formation of irregular filamentous structures.
authors: Unit of Virus Host Cell Interactions, UMR 5233 UJF-EMBL-CNRS, 6 rue Jules Horowitz, 38042 Grenoble Cedex 9, France.
authors: Francine C A
authors: FC
authors: Gerard
authors: Euripedes de Almeida
authors: Ede A
authors: Ribeiro
authors: Aurélie A V
authors: AA
authors: Albertini
authors: Irina
authors: I
authors: Gutsche
authors: Guiseppe
authors: G
authors: Zaccai
authors: Rob W H
authors: RW
authors: Ruigrok
authors: Marc
authors: M
authors: Jamin
mesh_terms: D002850:Chromatography, Gel; D019281:Dimerization; D007700:Kinetics; D008970:Molecular Weight; D033363:Neutron Diffraction; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D010766:Phosphorylation; D020836:Protein Structure, Quaternary; D012209:Rhabdoviridae / Q000737:chemistry*; D053838:Scattering, Small Angle; D012996:Solutions; D013696:Temperature
chemical_list: D010750:Phosphoproteins; D012996:Solutions