Recruitment of a nicotinic acetylcholine receptor mutant lacking cytoplasmic tyrosine residues in its beta subunit into agrin-induced aggregates.
During synaptogenesis at the vertebrate skeletal neuromuscular junction, acetylcholine receptors (AChRs) form high-density aggregates opposite the presynaptic terminal in response to nerve-derived agrin. Agrin has been shown to stimulate tyrosine phosphorylation of a muscle-specific receptor tyrosine kinase MuSK and of the AChR beta subunit, and tyrosine kinase inhibitors and a tyrosine kinase-deficient mutant of MuSK prevent AChR aggregation. To evaluate the role of tyrosine phosphorylation of the AChR beta subunit in receptor aggregation, we replaced all three putative cytoplasmic tyrosine residues of the AChR beta subunit with phenylalanine residues and expressed the mutant receptors in cultured myotubes. Upon agrin treatment, transfected myotubes formed AChR aggregates that contained receptors with mutant beta subunits. Thus, AChRs can be recruited into agrin-induced specializations by protein-protein interactions that do not depend on tyrosine phosphorylation of the AChR beta subunit.
Department of Physiology and Biophysics, C240, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, Colorado, 80262, USA.
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Meyer
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Wallace
D018171:Agrin / Q000502:physiology*; D000818:Animals; D000911:Antibodies, Monoclonal; D002642:Chick Embryo; D003593:Cytoplasm; D015870:Gene Expression / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000201:enzymology; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D010766:Phosphorylation; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D011950:Receptors, Cholinergic; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D013045:Species Specificity; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism*
D018171:Agrin; D000911:Antibodies, Monoclonal; D011950:Receptors, Cholinergic; D011978:Receptors, Nicotinic; D014443:Tyrosine; C095714:MUSK protein, human; D020794:Receptor Protein-Tyrosine Kinases