Opposing Roles of FoxP1 and Nfat3 in Transcriptional Control of Cardiomyocyte Hypertrophy

Abstract

Cardiac homeostasis is maintained by a balance of growth-promoting and growth-modulating factors. Sustained elevation of calcium signaling can induce cardiac hypertrophy through activation of Nfat family transcription factors. FoxP family transcription factors are known to interact with Nfat proteins and to modulate their transcriptional activities in lymphocytes. We investigated FoxP1 interaction with Nfat3 (Nfatc4) and their effects on transcription of hypertrophy-associated genes in neonatal rat cardiomyocytes. FoxP1-Nfat3 complexes were visualized using bimolecular fluorescence complementation (BiFC) analysis. Calcineurin activation induced FoxP1-Nfat3 BiFC complex formation. Amino acid substitutions in the predicted interaction interface inhibited it. FoxP1 repressed hypertrophy-associated genes (Myh7, Rcan1, Cx43, Anf, and Bnp) and counteracted their activation by constitutively nuclear Nfat3 (cnNfat3). In contrast, FoxP1 activated genes that maintain normal heart functions (Myh6 and p57Kip2) and cnNfat3 counteracted their activation by FoxP1. Amino acid substitutions in FoxP1 or cnNfat3 that inhibited their interaction abrogated the activation of hypertrophy-associated gene transcription by cnNfat3 and the repression of these genes by FoxP1. FoxP1 and Nfat3 co-occupied the promoter regions of hypertrophy-associated genes in neonatal and adult heart tissue. FoxP1 counteracted hypertrophic cardiomyocyte growth, and connexin 43 mislocalization caused by cnNfat3 expression. These data suggest that the opposing transcriptional activities of FoxP1 and Nfat3 maintain cardiomyocyte homeostasis.

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Retraction for Bai and Kerppola, “Opposing Roles of FoxP1 and Nfat3 in Transcriptional Control of Cardiomyocyte Hypertrophy”

ACKNOWLEDGMENTS

We thank Edward E. Morrisey (University of Pennsylvania) for anti-FoxP1 antiserum, the late Nancy Rice (NCI) for anti-Nfat3 antiserum, Gerald Thiel (University of Saarland Medical Center) for the CnA expression vector, Beverly A. Rothermel (UTSW) and R. Sanders Williams (Duke University) for Rcan1 reporters, Kenneth M Baldwin (University of California, Irvine) for Myh6 and Myh7 reporters, and Margaret Westfall and Ravi K. Birla for assistance with cardiomyocyte isolation and culture. We thank members of the Kerppola laboratory for helpful suggestions and criticisms.

This work was funded by NIDA (R01 DA030339) and by a grant from the Dana Foundation. S.B. received funding as a Center for Organogenesis Fellow.