title: Molecular basis for the interaction between rabies virus phosphoprotein P and the dynein light chain LC8: dissociation of dynein-binding properties and transcriptional functionality of P.
abstract: The lyssavirus phosphoprotein P is a co-factor of the viral RNA polymerase and plays a central role in virus transcription and replication. It has been shown previously that P interacts with the dynein light chain LC8, which is involved in minus end-directed movement of organelles along microtubules. Co-immunoprecipitation experiments and the two-hybrid system were used to map the LC8-binding site to the sequence (139)RSSEDKSTQTTGR(151). Site-directed mutagenesis of residues D(143) and Q(147) to an A residue abolished binding to LC8. The P-LC8 association is not required for virus transcription, since the double mutant was not affected in its transcription ability in a minigenome assay. Based on the crystal structure of LC8 bound to a peptide from neuronal nitric oxide synthase, a model for the complex between the peptide spanning residues 140-150 of P and LC8 is proposed. This model suggests that P binds LC8 in a manner similar to other LC8 cellular partners.
authors.affiliation: Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Poisson
authors.affiliation: Laboratoire des Lyssavirus, Institut Pasteur 25 rue du Dr Roux, 75724 Paris Cedex 15, France2.
authors.forename: Eleonore
authors.initials: E
authors.lastname: Real
authors.affiliation: Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
authors.forename: Yves
authors.initials: Y
authors.lastname: Gaudin
authors.affiliation: Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Vaney
authors.affiliation: Department of Biochemistry, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, Connecticut 06030-3305, USA3.
authors.forename: Stephen
authors.initials: S
authors.lastname: King
authors.affiliation: Laboratoire des Lyssavirus, Institut Pasteur 25 rue du Dr Roux, 75724 Paris Cedex 15, France2.
authors.forename: Yves
authors.initials: Y
authors.lastname: Jacob
authors.affiliation: Laboratoire des Lyssavirus, Institut Pasteur 25 rue du Dr Roux, 75724 Paris Cedex 15, France2.
authors.forename: Noël
authors.initials: N
authors.lastname: Tordo
authors.affiliation: Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
authors.forename: Danielle
authors.initials: D
authors.lastname: Blondel
mesh_terms: D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins; D004398:Dyneins; D008958:Models, Molecular; D018832:Molecular Chaperones; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D011820:Rabies virus / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques; D015678:Viral Structural Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D029721:Drosophila Proteins; D018832:Molecular Chaperones; C103961:P phosphoprotein, Rabies virus; D010750:Phosphoproteins; D015678:Viral Structural Proteins; D004398:Dyneins