STAT4 serine phosphorylation is critical for IL-12-induced IFN-γ production but not for cell proliferation

Akio Morinobu, Massimo Gadina, Warren Strober, Roberta Visconti, Albert Fornace, Cristina Montagna, Gerald M. Feldman, Ryuta Nishikomori, John J. O'Shea

Research output: Contribution to journalArticle

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Abstract

T helper 1 (TH1) differentiation and IFN-γ production are crucial in cell-mediated immune responses. IL-12 is an important regulator of this process and mediates its effects through signal transducer and activator of transcription 4 (STAT4). IFN-γ production is also regulated by the p38 mitogen-activated kinase pathway, although the mechanisms are ill-defined. We show here that GADD45-β and GADD45-γ can induce STAT4 S721 phosphorylation via the MKK6/p38 pathway. Thus, STAT4 could be a target that accounts for the defects in cell-mediated immunity associated with perturbations in the p38 pathway. To investigate the biological significance of STAT4 S721 phosphorylation, we reconstituted primary spleen cells from STAT4-deficient mice with wild-type and mutated STAT4, by using a retroviral gene transduction. We demonstrated that expression of wild-type STAT4, but not the S721A mutant, restored normal TH1 differentiation and IFN-γ synthesis. The inability of STAT4 S721 to restore IFN-γ production was not caused by decreased IL-12R expression because the STAT4 S721 mutant also failed to restore IFN-γ production in STAT4-deficient IL-12Rβ2 transgenic cells. Importantly, STAT4 S721A-transduced cells showed normal proliferative response to IL-12, illustrating that serine phosphorylation is not required for IL-12-induced proliferation. Additionally, the results imply the existence of STAT4 serine phosphorylation-dependent and -independent target genes. We conclude that phosphorylation of STAT4 on both tyrosine and serine residues is important in promoting normal TH1 differentiation and IFN-γ secretion.

Original languageEnglish (US)
Pages (from-to)12281-12286
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume99
Issue number19
DOIs
StatePublished - Sep 17 2002
Externally publishedYes

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STAT4 Transcription Factor
Interleukin-12
Serine
Phosphorylation
Cell Proliferation

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

STAT4 serine phosphorylation is critical for IL-12-induced IFN-γ production but not for cell proliferation. / Morinobu, Akio; Gadina, Massimo; Strober, Warren; Visconti, Roberta; Fornace, Albert; Montagna, Cristina; Feldman, Gerald M.; Nishikomori, Ryuta; O'Shea, John J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, No. 19, 17.09.2002, p. 12281-12286.

Research output: Contribution to journalArticle

Morinobu, Akio ; Gadina, Massimo ; Strober, Warren ; Visconti, Roberta ; Fornace, Albert ; Montagna, Cristina ; Feldman, Gerald M. ; Nishikomori, Ryuta ; O'Shea, John J. / STAT4 serine phosphorylation is critical for IL-12-induced IFN-γ production but not for cell proliferation. In: Proceedings of the National Academy of Sciences of the United States of America. 2002 ; Vol. 99, No. 19. pp. 12281-12286.
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T1 - STAT4 serine phosphorylation is critical for IL-12-induced IFN-γ production but not for cell proliferation

AU - Morinobu, Akio

AU - Gadina, Massimo

AU - Strober, Warren

AU - Visconti, Roberta

AU - Fornace, Albert

AU - Montagna, Cristina

AU - Feldman, Gerald M.

AU - Nishikomori, Ryuta

AU - O'Shea, John J.

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N2 - T helper 1 (TH1) differentiation and IFN-γ production are crucial in cell-mediated immune responses. IL-12 is an important regulator of this process and mediates its effects through signal transducer and activator of transcription 4 (STAT4). IFN-γ production is also regulated by the p38 mitogen-activated kinase pathway, although the mechanisms are ill-defined. We show here that GADD45-β and GADD45-γ can induce STAT4 S721 phosphorylation via the MKK6/p38 pathway. Thus, STAT4 could be a target that accounts for the defects in cell-mediated immunity associated with perturbations in the p38 pathway. To investigate the biological significance of STAT4 S721 phosphorylation, we reconstituted primary spleen cells from STAT4-deficient mice with wild-type and mutated STAT4, by using a retroviral gene transduction. We demonstrated that expression of wild-type STAT4, but not the S721A mutant, restored normal TH1 differentiation and IFN-γ synthesis. The inability of STAT4 S721 to restore IFN-γ production was not caused by decreased IL-12R expression because the STAT4 S721 mutant also failed to restore IFN-γ production in STAT4-deficient IL-12Rβ2 transgenic cells. Importantly, STAT4 S721A-transduced cells showed normal proliferative response to IL-12, illustrating that serine phosphorylation is not required for IL-12-induced proliferation. Additionally, the results imply the existence of STAT4 serine phosphorylation-dependent and -independent target genes. We conclude that phosphorylation of STAT4 on both tyrosine and serine residues is important in promoting normal TH1 differentiation and IFN-γ secretion.

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