An 83-nucleotide promoter of the acetylcholine receptor epsilon-subunit gene confers preferential synaptic expression in mouse muscle.
The expression of the acetylcholine receptor epsilon-subunit gene is restricted to the endplate of adult muscle fibers. We have started to study the regulatory elements of the epsilon-subunit gene promoter that are important for its synaptic expression. We used, for this purpose, a rapid method of in vivo expression after DNA injection into the muscle tissue [Wolff, J. A., Malone, R. W., Williams, P., Chong, W., Acsadi, G., Jani, A. & Felgner, P. L. (1990) Science 247, 1465-1468]. Our results show that a construction containing 83 nucleotides upstream from the transcription start site is sufficient to obtain preferential endplate expression. Moreover, mutation of a MyoD binding site located around position-70 does not alter this synaptic expression. We also studied the expression of this promoter in vitro in muscle primary cultures and showed the presence of a positive element between positions -122 and -83. Comparison of in vivo and in vitro results reveals that the elements important for in vivo localization at the synapse and in vitro expression in cultured muscle cells may differ.
Duclert
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A
Centre National de la Recherche Scientifique 0210, Département des Biotechnologies, Institut Pasteur, Paris, France.
Savatier
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Changeux
J P
JP
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D015698:Genomic Library; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D046911:Macromolecular Substances; D051379:Mice; D008811:Mice, Inbred DBA; D008969:Molecular Sequence Data; D009132:Muscles / Q000294:innervation / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D011950:Receptors, Cholinergic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion; D013569:Synapses / Q000502:physiology*; D014162:Transfection
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D011950:Receptors, Cholinergic; D011993:Recombinant Fusion Proteins; D004247:DNA; D008156:Luciferases