p204 protein overcomes the inhibition of the differentiation of P19 murine embryonal carcinoma cells to beating cardiac myocytes by Id proteins

doi:10.1074/jbc.M511748200

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Papers In Press, published online ahead of print March 22, 2006
J. Biol. Chem, 10.1074/jbc.M511748200
Submitted on October 31, 2005
Accepted on March 22, 2006

p204 protein overcomes the inhibition of the differentiation of P19 murine embryonal carcinoma cells to beating cardiac myocytes by Id proteins

Bo Ding, Chuan-ju Liu, Yan Huang, Jin Yu, Weihua Kong, and Peter Lengyel

MB&B, Yale University School of Medicine, New Haven, CT 06520-8024

Corresponding Author: peter.lengyel{at}yale.edu

We reported in an accompanying article that (i) the p204 protein is required for the differentiation of murine P19 embryonal carcinoma stem cells to beating cardiac myocytes, and (ii) the expression of p204 in the differentiating P19 cells is synergistically transactivated by the cardiac transcription factors Gata4, Nkx2.5 and Tbx5. Here we report that endogenous or ectopic Id (Inhibitor of differentiation) proteins inhibited the differentiation of P19 cells to myocytes. This was in consequence of the binding of Id1, Id2 or Id3 protein to the Gata4 and Nkx2.5 proteins and the resulting inhibition (i) of the binding of these transcription factors to each other and to DNA, as well as (ii) of their synergistic transactivation of the expression of various genes including atrial natriuretic factor and Ifi204 (encoding p204). p204 overcame this inhibition by Id proteins in consequence of (i) binding and sequestering Id proteins, (ii) accelerating their ubiquitination and degradation by proteasomes, and (iii) decreasing the level of Id proteins in the nucleus by increasing their translocation from the nucleus to the cytoplasm. (ii) and (iii) depended on the presence of the Nuclear Export Signal in p204. In the course of the differentiation Gata4, Nkx2.5 and p204 were components of a positive feedback loop. This arose in consequence of it that p204 overcame the inhibition of the synergistic activity of Gata4 and Nkx2.5 by the Id proteins.

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