PIM1 Kinase Phosphorylates the Human Transcription Factor FOXP3 at Serine 422 to Negatively Regulate Its Activity under Inflammation

• Published in: The Journal of biological chemistry 2014-09, Vol.289 (39), p.26872-26881
• Main Authors: Li, Zhiyuan, Lin, Fang, Zhuo, Changhua, Deng, Guoping, Chen, Zuojia, Yin, Shuying, Gao, Zhimei, Piccioni, Miranda, Tsun, Andy, Cai, Sanjun, Zheng, Song Guo, Zhang, Yu, Li, Bin
• Format: Article
• Language: English
• Subjects: Cell Proliferation - physiology; CTLA-4 Antigen - biosynthesis; CTLA-4 Antigen - genetics; CTLA-4 Antigen - immunology; Forkhead Transcription Factors - genetics; Forkhead Transcription Factors - immunology; Forkhead Transcription Factors - metabolism; Gene Expression Regulation - physiology; Gene Knockdown Techniques; Glucocorticoid-Induced TNFR-Related Protein - biosynthesis; Glucocorticoid-Induced TNFR-Related Protein - genetics; Glucocorticoid-Induced TNFR-Related Protein - immunology; HEK293 Cells; Humans; Immunology; Inflammation - genetics; Inflammation - immunology; Inflammation - metabolism; Interleukin-2 - biosynthesis; Interleukin-2 - genetics; Interleukin-2 - immunology; Interleukin-2 Receptor alpha Subunit - biosynthesis; Interleukin-2 Receptor alpha Subunit - genetics; Interleukin-2 Receptor alpha Subunit - immunology; Interleukin-6 - genetics; Interleukin-6 - immunology; Interleukin-6 - metabolism; Jurkat Cells; Phosphorylation - physiology; Proto-Oncogene Proteins c-pim-1 - genetics; Proto-Oncogene Proteins c-pim-1 - immunology; Proto-Oncogene Proteins c-pim-1 - metabolism; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - immunology; Receptors, Antigen, T-Cell - metabolism; Serine - genetics; Serine - immunology; Serine - metabolism; Signal Transduction - physiology; T-Lymphocytes, Regulatory - immunology; T-Lymphocytes, Regulatory - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M114.586651

Previous reports have suggested that human CD4+ CD25hiFOXP3+ T regulatory cells (Tregs) have functional plasticity and may differentiate into effector T cells under inflammation. The molecular mechanisms underlying these findings remain unclear. Here we identified the residue serine 422 of human FOXP3 as a phosphorylation site that regulates its function, which is not present in murine Foxp3. PIM1 kinase, which is highly expressed in human Tregs, was found to be able to interact with and to phosphorylate human FOXP3 at serine 422. T cell receptor (TCR) signaling inhibits PIM1 induction, whereas IL-6 promotes PIM1 expression in in vitro expanded human Tregs. PIM1 negatively regulates FOXP3 chromatin binding activity by specifically phosphorylating FOXP3 at Ser422. Our data also suggest that phosphorylation of FOXP3 at the Ser418 site could prevent FOXP3 phosphorylation at Ser422 mediated by PIM1. Knockdown of PIM1 in in vitro expanded human Tregs promoted FOXP3-induced target gene expression, including CD25, CTLA4, and glucocorticoid-induced tumor necrosis factor receptor (GITR), or weakened FOXP3-suppressed IL-2 gene expression and enhanced the immunosuppressive activity of Tregs. Furthermore, PIM1-specific inhibitor boosted FOXP3 DNA binding activity in in vitro expanded primary Tregs and also enhanced their suppressive activity toward the proliferation of T effector cells. Taken together, our findings suggest that PIM1 could be a new potential therapeutic target in the prevention and treatment of human-specific autoimmune diseases because of its ability to modulate the immunosuppressive activity of human Tregs.