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J Biol Chem, Vol. 273, Issue 52, 34904-34910, December 25, 1998
From the Department of Biochemistry, Medical Sciences Building,
University of Western Ontario, London, Ontario N6A 5C1, Canada
The Nkx2-5 homeodomain protein plays a key role
in cardiomyogenesis. Ectopic expression in frog and zebrafish embryos
results in an enlarged myocardium; however, expression of Nkx2-5 in
fibroblasts was not able to trigger the development of beating cardiac
muscle. In order to examine the ability of Nkx2-5 to modulate
endogenous cardiac specific gene expression in cells undergoing early
stages of differentiation, P19 cell lines overexpressing Nkx2-5 were differentiated in the absence of Me2SO. Nkx2-5
expression induced cardiomyogenesis in these cultures aggregated
without Me2SO. During differentiation into cardiac muscle,
Nkx2-5 expression resulted in the activation of myocyte enhancer
factor 2C (MEF2C), but not MEF2A, -B, or -D. In order to compare the
abilities of Nkx2-5 and MEF2C to induce cellular
differentiation, P19 cells overexpressing MEF2C were aggregated in the
absence of Me2SO. Similar to Nkx2-5, MEF2C
expression initiated cardiomyogenesis, resulting in the up-regulation of Brachyury T, bone morphogenetic protein-4, Nkx2-5, GATA-4, cardiac 
-actin, and myosin heavy chain expression. These findings indicate the presence of a positive regulatory network between
Nkx2-5 and MEF2C and show that both factors can direct early stages of
cell differentiation into a cardiomyogenic pathway.
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