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J. Biol. Chem., Vol. 275, Issue 42, 32398-32405, October 20, 2000
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From the Department of Biochemistry, Medical Sciences Building,
University of Western Ontario, London, Ontario N6A 5C1, Canada
The myogenic regulatory factors (MRFs), MyoD and
myogenin, can induce myogenesis in a variety of cell lines but not
efficiently in monolayer cultures of P19 embryonal carcinoma stem
cells. Aggregation of cells expressing MRFs, termed P19[MRF] cells,
results in an approximately 30-fold enhancement of myogenesis. Here we
examine molecular events occurring during P19 cell aggregation to
identify potential mechanisms regulating MRF activity. Although
myogenin protein was continually present in the nuclei of >90% of
P19[myogenin] cells, only a fraction of these cells differentiated.
Consequently, it appears that post-translational regulation controls
myogenin activity in a cell lineage-specific manner. A correlation was obtained between the expression of factors involved in somite patterning, including Wnt3a, Wnt5b, BMP-2/4, and Pax3, and the induction of myogenesis. Co-culturing P19[Wnt3a] cells with
P19[MRF] cells in monolayer resulted in a 5- to 8-fold increase in
myogenesis. Neither BMP-4 nor Pax3 was efficient in enhancing MRF
activity in unaggregated P19 cultures. Furthermore, BMP-4 abrogated the enhanced myogenesis induced by Wnt signaling. Consequently, signaling events resulting from Wnt3a expression but not BMP-4 signaling or Pax3
expression, regulate MRF function. Therefore, the P19 cell culture
system can be used to study the link between somite patterning events
and myogenesis.
Wnt Signaling Regulates the Function of MyoD and Myogenin*
,
*
This work was supported in part by a grant from the Medical
Research Council of Canada.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.
Supported by a Natural Sciences and Engineering Council of
Canada studentship and an Ontario Graduate Scholarship.
§
Supported by a Medical Research Council of Canada Scholarship
(development grant). To whom correspondence should be addressed: Dept.
of Biochemistry, Medical Sciences Bldg., University of Western Ontario,
London, Ontario N6A 5C1, Canada. Tel.: 519-661-2111 (ext. 86867); Fax:
519-661-3175; E-mail: skerjanc@julian.uwo.ca.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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