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Eur J Immunol ; 40(4): 1174-84, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20127678

ABSTRACT

The function of transcription factors can be critically regulated by SUMOylation. c-Maf, the cellular counterpart of v-maf oncogene, is a potent transactivator of the IL-4 gene in Th2 cells. We found in a yeast two-hybrid screen that c-Maf can interact with Ubc9 and PIAS1, two key enzymes of the SUMOylation pathway. In this study, we report that c-Maf co-localized with these two SUMO (small ubiquitin-like modifier) ligases in the nucleus and that c-Maf can be SUMOylated in vitro and also in primary Th2 cells. We also demonstrated that lysine-33 is the dominant, if not the only, SUMO acceptor site of c-Maf. SUMOylation of c-Maf attenuated its transcriptional activity. Reciprocally, a SUMOylation resistant c-Maf was more potent than WT-c-Maf in driving IL-4 production in c-Maf-deficient Th2 cells. Furthermore, we showed that ablation of the SUMO site did not alter the subcellular localization or the stability of c-Maf protein but instead enhanced its recruitment to the Il4-promoter. We conclude that SUMOylation at lysine-33 is a functionally critical post-translational modification event of c-Maf in Th cells.


Subject(s)
Interleukin-4/biosynthesis , Protein Inhibitors of Activated STAT/physiology , Protein Processing, Post-Translational , Proto-Oncogene Proteins c-maf/physiology , Small Ubiquitin-Related Modifier Proteins/physiology , Th2 Cells/metabolism , Ubiquitin-Conjugating Enzymes/physiology , Amino Acid Sequence , Animals , Cell Line , Cells, Cultured/metabolism , Humans , Interleukin-4/genetics , Kidney , Lysine/chemistry , Mice , Molecular Sequence Data , Protein Inhibitors of Activated STAT/chemistry , Protein Inhibitors of Activated STAT/isolation & purification , Protein Interaction Mapping , Proto-Oncogene Proteins c-maf/chemistry , Recombinant Fusion Proteins/physiology , Sequence Alignment , Sequence Homology, Amino Acid , Small Ubiquitin-Related Modifier Proteins/chemistry , Small Ubiquitin-Related Modifier Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/isolation & purification , Transcription, Genetic , Two-Hybrid System Techniques , Ubiquitin-Conjugating Enzymes/chemistry
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