Dual Role of Sumoylation in the Nuclear Localization and Transcriptional Activation of NFAT1

The nuclear import of nuclear factor of activated T cells (NFAT) transcription factors is critical for regulating NFAT activity. Here we demonstrate that the sumoylation of NFAT1 defines a novel mechanism of the nuclear anchorage and transcriptional activation downstream from the known mechanism of calcineurin-mediated dephosphorylation and nuclear import. We show that Lys 684 and Lys 897 of NFAT1 can be sumoylated. The sumoylation at Lys 684 is required for NFAT1 transcriptional activity and subsequent sumoylation of Lys 897 , whereas the sumoylation of Lys 897 is only required for nuclear anchorage. Because Lys 897 of NFAT1 is not conserved among other members of the NFAT family, we propose that sumoylation of Lys 897 may provide a mechanism for NFAT1 isotype-specific regulation of nuclear anchorage and transcriptional activation. Furthermore, we found that treatment with both ionomycin and phorbol 12-myristate 13-acetate ensured efficient nuclear anchorage with the recruitment of NFAT1 into the SUMO-1 bodies, whereas treatment with ionomycin alone induced nuclear translocation of NFAT1 but not recruitment into the SUMO-1 bodies. Our results suggest that the recruitment of NFAT1 into SUMO-1 bodies may be required for the progressive transcriptional activity of NFAT1 upon co-stimulation with ionomycin and phorbol 12-myristate 13-acetate, whereas anergic transcription stimulated by ionomycin alone may occur without recruitment into the SUMO-1 bodies.