Cytoplasmic dynein LC8 interacts with lyssavirus phosphoprotein.
Using a yeast two-hybrid human brain cDNA library screen, the cytoplasmic dynein light chain (LC8), a 10-kDa protein, was found to interact strongly with the phosphoprotein (P) of two lyssaviruses: rabies virus (genotype 1) and Mokola virus (genotype 3). The high degree of sequence divergence between these P proteins (only 46% amino acid identity) favors the hypothesis that this interaction is a common property shared by all lyssaviruses. The P protein-dynein LC8 interaction was confirmed by colocalization with laser confocal microscopy in infected cells and by coimmunoprecipitation. The dynein-interacting P protein domain was mapped to the 186 amino acid residues of the N-terminal half of the protein. Dynein LC8 is a component of both cytoplasmic dynein and myosin V, which are involved in a wide range of intracellular motile events, such as microtubule minus-end directed organelle transport in axon "retrograde transport" and actin-based vesicle transport, respectively. Our results provide support for a model of viral nucleocapsid axoplasmic transport. Furthermore, the role of LC8 in cellular mechanisms other than transport, e.g., inhibition of neuronal nitric oxide synthase, suggests that the P protein interactions could be involved in physiopathological mechanisms of rabies virus-induced pathogenesis.
Jacob
Y
Y
Laboratoire des Lyssavirus, Institut Pasteur, 75724 Paris Cedex 15, France. yjacob@pasteur.fr
Badrane
H
H
Ceccaldi
P E
PE
Tordo
N
N
D000818:Animals; D001921:Brain / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D003593:Cytoplasm / Q000502:physiology; D018076:DNA, Complementary; D029721:Drosophila Proteins; D004398:Dyneins; D015723:Gene Library; D006801:Humans; D018114:Lyssavirus / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D018832:Molecular Chaperones; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D011820:Rabies virus / Q000235:genetics / Q000378:metabolism; D018353:Rhabdoviridae Infections / Q000821:virology; D020798:Two-Hybrid System Techniques; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism
D002352:Carrier Proteins; D018076:DNA, Complementary; D029721:Drosophila Proteins; D018832:Molecular Chaperones; C103961:P phosphoprotein, Rabies virus; D010750:Phosphoproteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; D004398:Dyneins