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J. Biol. Chem., Vol. 276, Issue 20, 17603-17609, May 18, 2001
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From the Department of Cellular and Molecular Medicine and Centre
for Neuromuscular Disease, Faculty of Medicine, University of Ottawa,
Ottawa, Ontario, Canada K1H 8M5
In this study, we examined whether an intronic
N-box motif is involved in the expression of acetylcholinesterase
(AChE) during myogenesis. We determined that AChE transcripts are
barely detectable in cultured myoblasts and that their levels increase
dramatically in myotubes. Nuclear run-on assays revealed that this
increase was accompanied by a parallel induction in the transcriptional activity of the AChE gene. These changes in transcription
were also observed in transfection experiments using AChE
promoter-reporter gene constructs. Mutation of the intronic N-box at
position +755 base pairs (bp) reduced by more than 70% expression of
the reporter gene in myotubes. Disruption of an adjacent E-box, at
position +767 bp, also reduced expression of the reporter gene
following myogenic differentiation. Co-transfection experiments using
AChE promoter-reporter gene constructs and a myogenin expression vector showed that expression of this regulatory factor increased expression of the reporter gene in myotubes. Although the AChE promoter contains multiple E-boxes, mutation of this intronic one was sufficient to
prevent the myogenin-induced increase in reporter gene expression. Together, these results indicate that changes in AChE gene
transcription occur during myogenesis and highlight the contribution of
the intronic N- and E-box motifs in the developmental regulation of the
AChE gene in skeletal muscle.
Role of Intronic E- and N-box Motifs in the Transcriptional
Induction of the Acetylcholinesterase Gene during Myogenic
Differentiation*
,
*
This work was supported in part by operating grants from the
Canadian Institutes of Health Research (CIHR) (to B. J. J.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of the William T. McEachern Fellowship for Doctoral students.
§
An Investigator of the CIHR. To whom correspondence should be
addressed: Dept. of Cellular and Molecular Medicine, Faculty of
Medicine, University of Ottawa, 451 Smyth Rd., Ottawa, Ontario K1H 8M5,
Canada. Tel.: 613-562-5800 (ext. 8383); Fax: 613-562-5636; E-mail:
jasmin@uottawa.ca.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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