DOI: 10.1073/pnas.96.24.13956. Stress-induced phosphorylation of STAT1 at Ser727 requires p38 mitogen-activated protein kinase whereas IFN-γ uses a different signaling pathway

Stress-induced phosphorylation of STAT1 at Ser727 requires p38 mitogen-activated protein kinase whereas IFN-γ uses a different signaling pathway

10.1073/pnas.96.24.13956

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

STAT1 is an essential transcription factor for macrophage activation by IFN-γ and requires phosphorylation of the C-terminal Ser727 for transcriptional activity. In macrophages, Ser727 phosphorylation in response to bacterial lipopolysaccharide (LPS), UV irradiation, or TNF-α occurred through a signaling path sensitive to the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580 whereas IFN-γ-mediated Ser727 phosphorylation was not inhibited by the drug. Consistently, SB203580 did not affect IFN-γ-mediated, Stat1-dependent transcription but inhibited its enhancement by LPS. Furthermore, LPS, UV irradiation, and TNF-α caused activation of p38 MAPK whereas IFN-γ did not. An essential role for p38 MAPK activity in STAT1 Ser727 phosphorylation was confirmed by using cells expressing an SB203580-resistant p38 MAPK. In such cells, STAT1 Ser727 phosphorylation in response to UV irradiation was found to be SB203580 insensitive. Targeted disruption of themapkap-k2gene, encoding a kinase downstream of p38 MAPK with a key role in LPS-stimulated TNF-α production and stress-induced heat shock protein 25 phosphorylation, was without a significant effect on UV-mediated Ser727 phosphorylation. The recombinant Stat1 C terminus was phosphorylatedin vitroby p38MAPKα and β but not by MAPK-activated protein kinase 2. Janus kinase 2 activity, previously reported to be required for IFN-γ-mediated Ser727 phosphorylation, was not needed for LPS-mediated Ser727 phosphorylation, and activation of Janus kinase 2 did not cause the appearance of STAT1 Ser727 kinase activity. Our data suggest that STAT1 is phosphorylated at Ser727 by a stress-activated signaling pathway either through p38 MAPK directly or through an unidentified kinase downstream of p38MAPK.

Bibliography

Kovarik, P., Stoiber, D., Eyers, P. A., Menghini, R., Neininger, A., Gaestel, M., Cohen, P., & Decker, T. (1999). Stress-induced phosphorylation of STAT1 at Ser727 requires p38 mitogen-activated protein kinase whereas IFN-Î³ uses a different signaling pathway. Proceedings of the National Academy of Sciences, 96(24), 13956â13961.

Authors 8

Pavel Kovarik (first)
Dagmar Stoiber (additional)
Patrick A. Eyers (additional)
Rossella Menghini (additional)
Armin Neininger (additional)
Matthias Gaestel (additional)
Philip Cohen (additional)
Thomas Decker (additional)

References 41 Referenced 226

10.1126/science.8197455
10.1146/annurev.immunol.16.1.293
10.1016/S0092-8674(00)81289-1
10.1016/S0092-8674(00)81288-X
10.1089/jir.1997.17.121
10.1016/0092-8674(95)90311-9
10.1073/pnas.93.15.7673
10.1128/jvi.70.1.647-650.1996
10.1093/emboj/17.13.3660
10.1096/fasebj.9.9.7601337
10.1007/s001090050063
10.1074/jbc.271.40.24313
10.1038/334715a0
10.1128/MCB.15.8.4353
10.1016/0092-8674(94)90277-1
10.1016/0092-8674(94)90278-X
10.1074/jbc.271.14.8488
10.1016/0014-5793(96)00816-2
10.1093/emboj/17.12.3372
10.1093/emboj/16.8.1909
10.1093/emboj/16.8.1921
10.1093/emboj/17.15.4426
10.1074/jbc.270.46.27489
10.1016/0014-5793(95)00357-F
10.1002/jcp.1041300316
10.1016/S0014-5793(99)00552-9
10.1038/10061
10.1016/S1074-5521(98)90170-3
10.1074/jbc.270.45.27213
J Sambrook, E F Fritsch, T Maniatis Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Lab. Press, Plainview, NY, 1989). / Molecular Cloning: A Laboratory Manual by Sambrook J (1989)
10.1073/pnas.93.7.2774
K K Hale, D Trollinger, M Rihanek, C L Manthey J Immunol 162, 4246–4252 (1999). (10.4049/jimmunol.162.7.4246) / J Immunol by Hale K K (1999)
10.1016/S0960-9822(02)00649-8
10.1128/MCB.17.11.6618
T Reimann, D Buscher, R A Hipskind, S Krautwald, M L Lohmann Matthes, M Baccarini J Immunol 153, 5740–5749 (1994). (10.4049/jimmunol.153.12.5740) / J Immunol by Reimann T (1994)
10.1016/S1074-5521(97)90194-0
10.1038/nsb0497-311
10.1093/emboj/17.12.3363
J A Gollob, C P Schnipper, E A Murphy, J Ritz, D A Frank J Immunol 162, 4472–4481 (1999). (10.4049/jimmunol.162.8.4472) / J Immunol by Gollob J A (1999)
10.1093/emboj/18.9.2480
10.1093/emboj/18.20.5601

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:40 a.m.)
Deposited	2 years, 4 months ago (April 22, 2023, 8:26 a.m.)
Indexed	1 month, 2 weeks ago (July 19, 2025, 11:53 p.m.)
Issued	25 years, 9 months ago (Nov. 23, 1999)
Published	25 years, 9 months ago (Nov. 23, 1999)
Published Online	25 years, 9 months ago (Nov. 23, 1999)
Published Print	25 years, 9 months ago (Nov. 23, 1999)

Funders 0

None

BibTeX

@article{Kovarik_1999, title={Stress-induced phosphorylation of STAT1 at Ser727 requires p38 mitogen-activated protein kinase whereas IFN-γ uses a different signaling pathway}, volume={96}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.96.24.13956}, DOI={10.1073/pnas.96.24.13956}, number={24}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Kovarik, Pavel and Stoiber, Dagmar and Eyers, Patrick A. and Menghini, Rossella and Neininger, Armin and Gaestel, Matthias and Cohen, Philip and Decker, Thomas}, year={1999}, month=nov, pages={13956–13961} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        20
      ]
    ],
    "date-time": "2025-07-20T03:53:35Z",
    "timestamp": 1752983615224
  },
  "reference-count": 41,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "24",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        1999,
        11,
        23
      ]
    ]
  },
  "abstract": "<jats:p>STAT1 is an essential transcription factor for macrophage activation by IFN-\u03b3 and requires phosphorylation of the C-terminal Ser727 for transcriptional activity. In macrophages, Ser727 phosphorylation in response to bacterial lipopolysaccharide (LPS), UV irradiation, or TNF-\u03b1 occurred through a signaling path sensitive to the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580 whereas IFN-\u03b3-mediated Ser727 phosphorylation was not inhibited by the drug. Consistently, SB203580 did not affect IFN-\u03b3-mediated, Stat1-dependent transcription but inhibited its enhancement by LPS. Furthermore, LPS, UV irradiation, and TNF-\u03b1 caused activation of p38 MAPK whereas IFN-\u03b3 did not. An essential role for p38 MAPK activity in STAT1 Ser727 phosphorylation was confirmed by using cells expressing an SB203580-resistant p38 MAPK. In such cells, STAT1 Ser727 phosphorylation in response to UV irradiation was found to be SB203580 insensitive. Targeted disruption of the<jats:italic>mapkap-k2</jats:italic>gene, encoding a kinase downstream of p38 MAPK with a key role in LPS-stimulated TNF-\u03b1 production and stress-induced heat shock protein 25 phosphorylation, was without a significant effect on UV-mediated Ser727 phosphorylation. The recombinant Stat1 C terminus was phosphorylated<jats:italic>in vitro</jats:italic>by p38MAPK\u03b1 and \u03b2 but not by MAPK-activated protein kinase 2. Janus kinase 2 activity, previously reported to be required for IFN-\u03b3-mediated Ser727 phosphorylation, was not needed for LPS-mediated Ser727 phosphorylation, and activation of Janus kinase 2 did not cause the appearance of STAT1 Ser727 kinase activity. Our data suggest that STAT1 is phosphorylated at Ser727 by a stress-activated signaling pathway either through p38 MAPK directly or through an unidentified kinase downstream of p38MAPK.</jats:p>",
  "DOI": "10.1073/pnas.96.24.13956",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T14:40:52Z",
    "timestamp": 1027694452000
  },
  "page": "13956-13961",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 226,
  "title": "Stress-induced phosphorylation of STAT1 at Ser727 requires p38 mitogen-activated protein kinase whereas IFN-\u03b3 uses a different signaling pathway",
  "prefix": "10.1073",
  "volume": "96",
  "author": [
    {
      "given": "Pavel",
      "family": "Kovarik",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    },
    {
      "given": "Dagmar",
      "family": "Stoiber",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    },
    {
      "given": "Patrick A.",
      "family": "Eyers",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    },
    {
      "given": "Rossella",
      "family": "Menghini",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    },
    {
      "given": "Armin",
      "family": "Neininger",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    },
    {
      "given": "Matthias",
      "family": "Gaestel",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    },
    {
      "given": "Philip",
      "family": "Cohen",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    },
    {
      "given": "Thomas",
      "family": "Decker",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Vienna Biocenter, Institute of Microbiology and Genetics, Dr. Bohr-Gasse 9, A-1030 Vienna, Austria; MRC Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, DD1 4HN Scotland, United Kingdom; and Martin-Luther-University Halle-Wittenberg, Innovationskolleg Zellspezialisierung, Hoher Weg 8, 06120 Halle, Germany"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        1999,
        11,
        23
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_4_1_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.8197455"
    },
    {
      "key": "e_1_3_4_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.immunol.16.1.293"
    },
    {
      "key": "e_1_3_4_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0092-8674(00)81289-1"
    },
    {
      "key": "e_1_3_4_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0092-8674(00)81288-X"
    },
    {
      "key": "e_1_3_4_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1089/jir.1997.17.121"
    },
    {
      "key": "e_1_3_4_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(95)90311-9"
    },
    {
      "key": "e_1_3_4_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.93.15.7673"
    },
    {
      "key": "e_1_3_4_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/jvi.70.1.647-650.1996"
    },
    {
      "key": "e_1_3_4_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/17.13.3660"
    },
    {
      "key": "e_1_3_4_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1096/fasebj.9.9.7601337"
    },
    {
      "key": "e_1_3_4_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/s001090050063"
    },
    {
      "key": "e_1_3_4_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.40.24313"
    },
    {
      "key": "e_1_3_4_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/334715a0"
    },
    {
      "key": "e_1_3_4_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MCB.15.8.4353"
    },
    {
      "key": "e_1_3_4_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(94)90277-1"
    },
    {
      "key": "e_1_3_4_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(94)90278-X"
    },
    {
      "key": "e_1_3_4_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.14.8488"
    },
    {
      "key": "e_1_3_4_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0014-5793(96)00816-2"
    },
    {
      "key": "e_1_3_4_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/17.12.3372"
    },
    {
      "key": "e_1_3_4_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/16.8.1909"
    },
    {
      "key": "e_1_3_4_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/16.8.1921"
    },
    {
      "key": "e_1_3_4_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/17.15.4426"
    },
    {
      "key": "e_1_3_4_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.46.27489"
    },
    {
      "key": "e_1_3_4_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0014-5793(95)00357-F"
    },
    {
      "key": "e_1_3_4_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/jcp.1041300316"
    },
    {
      "key": "e_1_3_4_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0014-5793(99)00552-9"
    },
    {
      "key": "e_1_3_4_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/10061"
    },
    {
      "key": "e_1_3_4_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1074-5521(98)90170-3"
    },
    {
      "key": "e_1_3_4_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.45.27213"
    },
    {
      "key": "e_1_3_4_30_2",
      "volume-title": "Molecular Cloning: A Laboratory Manual",
      "author": "Sambrook J",
      "year": "1989",
      "unstructured": "J Sambrook, E F Fritsch, T Maniatis Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Lab. Press, Plainview, NY, 1989)."
    },
    {
      "key": "e_1_3_4_31_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.93.7.2774"
    },
    {
      "key": "e_1_3_4_32_2",
      "doi-asserted-by": "crossref",
      "first-page": "4246",
      "DOI": "10.4049/jimmunol.162.7.4246",
      "volume": "162",
      "author": "Hale K K",
      "year": "1999",
      "unstructured": "K K Hale, D Trollinger, M Rihanek, C L Manthey J Immunol 162, 4246\u20134252 (1999).",
      "journal-title": "J Immunol"
    },
    {
      "key": "e_1_3_4_33_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0960-9822(02)00649-8"
    },
    {
      "key": "e_1_3_4_34_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MCB.17.11.6618"
    },
    {
      "key": "e_1_3_4_35_2",
      "doi-asserted-by": "crossref",
      "first-page": "5740",
      "DOI": "10.4049/jimmunol.153.12.5740",
      "volume": "153",
      "author": "Reimann T",
      "year": "1994",
      "unstructured": "T Reimann, D Buscher, R A Hipskind, S Krautwald, M L Lohmann Matthes, M Baccarini J Immunol 153, 5740\u20135749 (1994).",
      "journal-title": "J Immunol"
    },
    {
      "key": "e_1_3_4_36_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1074-5521(97)90194-0"
    },
    {
      "key": "e_1_3_4_37_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/nsb0497-311"
    },
    {
      "key": "e_1_3_4_38_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/17.12.3363"
    },
    {
      "key": "e_1_3_4_39_2",
      "doi-asserted-by": "crossref",
      "first-page": "4472",
      "DOI": "10.4049/jimmunol.162.8.4472",
      "volume": "162",
      "author": "Gollob J A",
      "year": "1999",
      "unstructured": "J A Gollob, C P Schnipper, E A Murphy, J Ritz, D A Frank J Immunol 162, 4472\u20134481 (1999).",
      "journal-title": "J Immunol"
    },
    {
      "key": "e_1_3_4_40_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/18.9.2480"
    },
    {
      "key": "e_1_3_4_41_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/18.20.5601"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.96.24.13956",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        4,
        22
      ]
    ],
    "date-time": "2023-04-22T12:26:37Z",
    "timestamp": 1682166397000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.96.24.13956"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1999,
        11,
        23
      ]
    ]
  },
  "references-count": 41,
  "journal-issue": {
    "issue": "24",
    "published-print": {
      "date-parts": [
        [
          1999,
          11,
          23
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.96.24.13956"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.96.24.13956",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        1999,
        11,
        23
      ]
    ]
  },
  "assertion": [
    {
      "value": "1999-08-10",
      "order": 0,
      "name": "received",
      "label": "Received",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "1999-10-06",
      "order": 1,
      "name": "accepted",
      "label": "Accepted",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "1999-11-23",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}