Translational regulation of acetylcholinesterase by the RNA-binding protein pumilio-2 at the neuromuscular synapse

Emilio Marrero, Susana G Rossi, Andrew Darr, Pantelis Tsoulfas, Richard L Rotundo

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Acetylcholinesterase (AChE) is highly expressed at sites of nerve-muscle contact where it is regulated at both the transcriptional and post-transcriptional levels. Our understanding of the molecular mechanisms underlying its regulation is incomplete, but they appear to involve both translational and post-translational events as well. Here, we show that Pumilio-2 (PUM2), an RNA binding translational repressor, is highly localized at the neuromuscular junction where AChE mRNA concentrates. Immunoprecipitation of muscle cell extracts with a PUM2 specific antibody precipitated AChE mRNA, suggesting that PUM2 binds to the AChE transcripts in a complex. Gel shift assays using a bacterially expressed PUM2 RNA binding domain showed specific binding using wild type AChE 3′-UTR RNA segment that was abrogated by mutation of the consensus recognition site. Transfecting skeletal muscle cells with shRNAs specific for PUM2 up-regulated AChE expression, whereas overexpression of PUM2 decreased AChE activity. We conclude that PUM2 binds to AChE mRNA and regulates AChE expression translationally at the neuromuscular synapse. Finally, we found that PUM2 is regulated by the motor nerve suggesting a trans-synaptic mechanism for locally regulating translation of specific proteins involved in modulating synaptic transmission, analogous to CNS synapses.

Original languageEnglish
Pages (from-to)36492-36499
Number of pages8
JournalJournal of Biological Chemistry
Volume286
Issue number42
DOIs
StatePublished - Oct 21 2011

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RNA-Binding Proteins
Acetylcholinesterase
Synapses
Muscle
RNA
Muscle Cells
Messenger RNA
Cells
Neuromuscular Junction
Protein Biosynthesis
3' Untranslated Regions
Cell Extracts
Immunoprecipitation
Synaptic Transmission
Assays
Skeletal Muscle
Gels
Muscles
Mutation
Antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Translational regulation of acetylcholinesterase by the RNA-binding protein pumilio-2 at the neuromuscular synapse. / Marrero, Emilio; Rossi, Susana G; Darr, Andrew; Tsoulfas, Pantelis; Rotundo, Richard L.

In: Journal of Biological Chemistry, Vol. 286, No. 42, 21.10.2011, p. 36492-36499.

Research output: Contribution to journalArticle

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