Regulation of muscle acetylcholine receptor turnover by β subunit tyrosine phosphorylation

John B. Rudell, Michael J Ferns

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

At the neuromuscular junction (NMJ), the postsynaptic localization of muscle acetylcholine receptor (AChR) is regulated by neural signals and occurs via several processes including metabolic stabilization of the receptor. However, the molecular mechanisms that influence receptor stability remain poorly defined. Here, we show that neural agrin and the tyrosine phosphatase inhibitor, pervanadate slow the degradation of surface receptor in cultured muscle cells. Their action is mediated by tyrosine phosphorylation of the AChR β subunit, as agrin and pervandate had no effect on receptor half-life in AChR-β3F/3F muscle cells, which have targeted mutations of the β subunit cytoplasmic tyrosines. Moreover, in wild type AChR-β3Y muscle cells, we found a linear relationship between average receptor half-life and the percentage of AChR with phosphorylated β subunit, with half-lives of 12.7 and 23 h for nonphosphorylated and phosphorylated receptor, respectively. Surprisingly, pervanadate increased receptor half-life in AChR-β3Y myotubes in the absence of clustering, and agrin failed to increase receptor half-life in AChR-β3F/3F myotubes even in the presence of clustering. The metabolic stabilization of the AChR was mediated specifically by phosphorylation of βY390 as mutation of this residue abolished β subunit phosphorylation but did not affect δ subunit phosphorylation. Receptor stabilization also led to higher receptor levels, as agrin increased surface AChR by 30% in AChR-β3Y but not AChR-β3F/3F myotubes. Together, these findings identify an unexpected role for agrin-induced phosphorylation of βY390 in downregulating AChR turnover. This likely stabilizes AChR at developing synapses, and contributes to the extended half-life of AChR at adult NMJs.

Original languageEnglish (US)
Pages (from-to)399-410
Number of pages12
JournalDevelopmental Neurobiology
Volume73
Issue number5
DOIs
StatePublished - May 2013

Keywords

  • Acetylcholine receptor
  • Agrin
  • Neuromuscular junction
  • Phosphorylation
  • Synaptogenesis

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Developmental Neuroscience

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