Nuclear Factor YY1 Inhibits Transforming Growth Factor β- and Bone Morphogenetic Protein-Induced Cell Differentiation

Abstract

Smad proteins transduce transforming growth factor β (TGF-β) and bone morphogenetic protein (BMP) signals that regulate cell growth and differentiation. We have identified YY1, a transcription factor that positively or negatively regulates transcription of many genes, as a novel Smad-interacting protein. YY1 represses the induction of immediate-early genes to TGF-β and BMP, such as the plasminogen activator inhibitor 1 gene (PAI-1) and the inhibitor of differentiation/inhibitor of DNA binding 1 gene (Id-1). YY1 inhibits binding of Smads to their cognate DNA elements in vitro and blocks Smad recruitment to the Smad-binding element-rich region of the PAI-1 promoter in vivo. YY1 interacts with the conserved N-terminal Mad homology 1 domain of Smad4 and to a lesser extent with Smad1, Smad2, and Smad3. The YY1 zinc finger domain mediates the association with Smads and is necessary for the repressive effect of YY1 on Smad transcriptional activity. Moreover, downregulation of endogenous YY1 by antisense and small interfering RNA strategies results in enhanced transcriptional responses to TGF-β or BMP. Ectopic expression of YY1 inhibits, while knockdown of endogenous YY1 enhances, TGF-β- and BMP-induced cell differentiation. In contrast, overexpression or knockdown of YY1 does not affect growth inhibition induced by TGF-β or BMP. Accordingly, YY1 does not interfere with the regulation of immediate-early genes involved in the TGF-β growth-inhibitory response, the cell cycle inhibitors p15 and p21, and the proto-oncogene c-myc. In conclusion, YY1 represses Smad transcriptional activities in a gene-specific manner and thus regulates cell differentiation induced by TGF-β superfamily pathways.

ACKNOWLEDGMENTS

We thank O. Korchynskyi for stimulating discussions and for performing the experiments for Fig. 1F and T. Imamura for advice on the DNAP assays. We thank Santa Cruz Biotechnology, K. Sampath, N. Ferrara, F. M. Hoffman, A. Comer, S. Itoh, B. Lüscher, M. Austen, T. D. Gelehrter, X.-F. Wang, G. Mavrothalassitis, J.-M. Gauthier, D. Rifkin, K. Miyazono, B. Vogelstein, and N. Fusenig for valuable reagents.

This research was funded in part by grants from the Human Frontier Science Program (to A.M.) and the Dutch Cancer Society (to P.T.D.) (NKI 2000-2217) and by a STINT grant from the Swedish Foundation for International Cooperation in Research and Higher Education (to C.-H.H.). U.V. was supported by a postdoctoral fellowship from the French Association pour la Recherche sur le Cancer.

K. Kurisaki, A. Kurisaki, and U. Valcourt contributed equally to this work.