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.
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.
Poisson
Nicolas
N
Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
Real
Eleonore
E
Laboratoire des Lyssavirus, Institut Pasteur 25 rue du Dr Roux, 75724 Paris Cedex 15, France2.
Gaudin
Yves
Y
Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
Vaney
Marie-Christine
MC
Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
King
Stephen
S
Department of Biochemistry, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, Connecticut 06030-3305, USA3.
Jacob
Yves
Y
Laboratoire des Lyssavirus, Institut Pasteur 25 rue du Dr Roux, 75724 Paris Cedex 15, France2.
Tordo
Noël
N
Laboratoire des Lyssavirus, Institut Pasteur 25 rue du Dr Roux, 75724 Paris Cedex 15, France2.
Blondel
Danielle
D
Laboratoire de Génétique des Virus, CNRS, 91198 Gif sur Yvette, France1.
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*
D002352:Carrier Proteins; D029721:Drosophila Proteins; D018832:Molecular Chaperones; C103961:P phosphoprotein, Rabies virus; D010750:Phosphoproteins; D015678:Viral Structural Proteins; D004398:Dyneins