Association of muscle-specific kinase MuSK with the acetylcholine receptor in mammalian muscle.
During synaptogenesis at the neuromuscular junction, a neurally released factor, agrin, causes the clustering of acetylcholine receptors (AChRs) in the muscle membrane beneath the nerve terminal. Agrin acts through a specific receptor which is thought to have a receptor tyrosine kinase, MuSK, as one of its components. In agrin-treated muscle cells, both MuSK and the AChR become tyrosine phosphorylated. To determine how the activation of MuSK leads to AChR clustering, we have investigated their interaction in cultured C2 myotubes. Immunoprecipitation experiments showed that MuSK is associated with the AChR and that this association is increased by agrin treatment. Agrin also caused a transient activation of the AChR-associated MuSK, as demonstrated by MuSK phosphorylation. In agrin-treated myotubes, MuSK phosphorylation increased with the same time course as phosphorylation of the beta subunit of the AChR, but declined more quickly. Although both herbimycin and staurosporine blocked agrin-induced AChR phosphorylation, only herbimycin inhibited the phosphorylation of MuSK. These results suggest that although agrin increases the amount of activated MuSK that is associated with the AChR, MuSK is not directly responsible for AChR phosphorylation but acts through other kinases.
Fuhrer
C
C
Section on Synaptic Mechanisms, Laboratory of Cellular and Molecular Regulation, National Institute of Mental Health, NIH, Bethesda, MD 20892, USA.
Sugiyama
J E
JE
Taylor
R G
RG
Hall
Z W
ZW
D018171:Agrin / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D016227:Benzoquinones; D015153:Blotting, Western; D002038:Bungarotoxins / Q000378:metabolism; D002478:Cells, Cultured; D004789:Enzyme Activation; D006801:Humans; D047029:Lactams, Macrocyclic; D051379:Mice; D008954:Models, Biological; D009132:Muscles / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000737:chemistry / Q000378:metabolism*; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction / Q000502:physiology; D019311:Staurosporine / Q000494:pharmacology; D014162:Transfection
D018171:Agrin; D016227:Benzoquinones; D002038:Bungarotoxins; D047029:Lactams, Macrocyclic; D011809:Quinones; D011950:Receptors, Cholinergic; D017828:Rifabutin; D019000:Phosphotyrosine; C020754:herbimycin; C095714:MUSK protein, human; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; D019311:Staurosporine