IRAK1-independent pathways required for the interleukin-1-stimulated activation of the Tpl2 catalytic subunit and its dissociation from ABIN2

Hosea Handoyo, Margaret J. Stafford, Eamon McManus, Dionissios Baltzis, Mark Peggie, Philip Cohen

    Research output: Contribution to journalArticle

    18 Citations (Scopus)

    Abstract

    The protein kinase Tpl2 (tumour progression locus 2) is activated by LPS (lipopolysaccharide), TNF alpha (tumour necrosis factor alpha) and IL (interleukin)-1. Activation of the native Tpl2 complex by these agonists requires the IKK beta {I kappa B [inhibitor of NF-kappa B (nuclear factor kappa B)] kinase beta}-catalysed phosphorylation of the p105/NF-kappa B1 subunit and is accompanied by the release of the catalytic subunit from both p105/NF-kappa B1 and another subunit ABIN2 (A20-binding inhibitor of NF-kappa B2). In the present study we report that IL-1 activates the transfected Tpl2 catalytic subunit in an HEK (human embryonic kidney)-293 cell line that stably expresses the IL-1R (IL-1 receptor), but does not express the protein kinase IRAK1 (IL-1R-associated kinase). In these cells IL-1 does not activate IKK beta or induce the phosphorylation of p105/NF-kappa B1, and nor does the IKK beta inhibitor PSI 145 prevent the IL-1-induced activation of transfected Tpl2. However, the IL-1-stimulated activation of transfected Tpl2 in IRAK1-null cells or activation of the endogenous Tpl2 complex in IRAK1-expressing cells is suppressed by the protein kinase inhibitor PP2 by a mechanism that does not involve inhibition of Src family protein tyrosine kinases. The IL-1-stimulated activation of transfected Tpl2 is accompanied by its phosphorylation at Thr(290) and Ser(400) and by enhanced phosphorylation of Ser(62), which we demonstrate are autophosphorylation events catalysed by Tpl2 itself. We further show that IL-1 triggers the dissociation of Tpl2 from co-transfected ABIN2 in IRAK1-null IL-1R cells, which is not Suppressed by PP2 or by the inhibition of Tpl2 or IKK beta. These studies identify two new signalling events involved in activation of the native Tpl2 complex by IL-1. First, the IRAK1-, IKK beta- and PP2-independent dissociation of Tpl2 from ABIN2; secondly, the IRAK1- and IKK beta-independent, but PP2-sensitive, activation of the Tpl2 catalytic subunit.

    Original languageEnglish
    Pages (from-to)109-118
    Number of pages10
    JournalBiochemical Journal
    Volume424
    DOIs
    Publication statusPublished - 15 Nov 2009

    Keywords

    • innate immunity
    • interleukin-1 (IL-1)
    • interleukin-1-receptor-associated kinase 1 (IRAK1)
    • nuclear factor kappa B (NF-kappa B)
    • tumour necrosis factor (TNF)
    • tumour progression locus 2 (Tpl2)
    • KAPPA-B-KINASE
    • SIGNAL-REGULATED KINASE
    • PROTEIN-KINASE
    • MAP KINASE
    • LIPOPOLYSACCHARIDE ACTIVATION
    • NF-KAPPA-B1 P105
    • TERNARY COMPLEX
    • TNF-ALPHA
    • PHOSPHORYLATION
    • MACROPHAGES

    Cite this

    Handoyo, Hosea ; Stafford, Margaret J. ; McManus, Eamon ; Baltzis, Dionissios ; Peggie, Mark ; Cohen, Philip. / IRAK1-independent pathways required for the interleukin-1-stimulated activation of the Tpl2 catalytic subunit and its dissociation from ABIN2. In: Biochemical Journal. 2009 ; Vol. 424. pp. 109-118.
    @article{df6ff189e6a6438c8356c9505021e26f,
    title = "IRAK1-independent pathways required for the interleukin-1-stimulated activation of the Tpl2 catalytic subunit and its dissociation from ABIN2",
    abstract = "The protein kinase Tpl2 (tumour progression locus 2) is activated by LPS (lipopolysaccharide), TNF alpha (tumour necrosis factor alpha) and IL (interleukin)-1. Activation of the native Tpl2 complex by these agonists requires the IKK beta {I kappa B [inhibitor of NF-kappa B (nuclear factor kappa B)] kinase beta}-catalysed phosphorylation of the p105/NF-kappa B1 subunit and is accompanied by the release of the catalytic subunit from both p105/NF-kappa B1 and another subunit ABIN2 (A20-binding inhibitor of NF-kappa B2). In the present study we report that IL-1 activates the transfected Tpl2 catalytic subunit in an HEK (human embryonic kidney)-293 cell line that stably expresses the IL-1R (IL-1 receptor), but does not express the protein kinase IRAK1 (IL-1R-associated kinase). In these cells IL-1 does not activate IKK beta or induce the phosphorylation of p105/NF-kappa B1, and nor does the IKK beta inhibitor PSI 145 prevent the IL-1-induced activation of transfected Tpl2. However, the IL-1-stimulated activation of transfected Tpl2 in IRAK1-null cells or activation of the endogenous Tpl2 complex in IRAK1-expressing cells is suppressed by the protein kinase inhibitor PP2 by a mechanism that does not involve inhibition of Src family protein tyrosine kinases. The IL-1-stimulated activation of transfected Tpl2 is accompanied by its phosphorylation at Thr(290) and Ser(400) and by enhanced phosphorylation of Ser(62), which we demonstrate are autophosphorylation events catalysed by Tpl2 itself. We further show that IL-1 triggers the dissociation of Tpl2 from co-transfected ABIN2 in IRAK1-null IL-1R cells, which is not Suppressed by PP2 or by the inhibition of Tpl2 or IKK beta. These studies identify two new signalling events involved in activation of the native Tpl2 complex by IL-1. First, the IRAK1-, IKK beta- and PP2-independent dissociation of Tpl2 from ABIN2; secondly, the IRAK1- and IKK beta-independent, but PP2-sensitive, activation of the Tpl2 catalytic subunit.",
    keywords = "innate immunity, interleukin-1 (IL-1), interleukin-1-receptor-associated kinase 1 (IRAK1), nuclear factor kappa B (NF-kappa B), tumour necrosis factor (TNF), tumour progression locus 2 (Tpl2), KAPPA-B-KINASE, SIGNAL-REGULATED KINASE, PROTEIN-KINASE, MAP KINASE, LIPOPOLYSACCHARIDE ACTIVATION, NF-KAPPA-B1 P105, TERNARY COMPLEX, TNF-ALPHA, PHOSPHORYLATION, MACROPHAGES",
    author = "Hosea Handoyo and Stafford, {Margaret J.} and Eamon McManus and Dionissios Baltzis and Mark Peggie and Philip Cohen",
    year = "2009",
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    IRAK1-independent pathways required for the interleukin-1-stimulated activation of the Tpl2 catalytic subunit and its dissociation from ABIN2. / Handoyo, Hosea; Stafford, Margaret J.; McManus, Eamon; Baltzis, Dionissios; Peggie, Mark; Cohen, Philip.

    In: Biochemical Journal, Vol. 424, 15.11.2009, p. 109-118.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - IRAK1-independent pathways required for the interleukin-1-stimulated activation of the Tpl2 catalytic subunit and its dissociation from ABIN2

    AU - Handoyo, Hosea

    AU - Stafford, Margaret J.

    AU - McManus, Eamon

    AU - Baltzis, Dionissios

    AU - Peggie, Mark

    AU - Cohen, Philip

    PY - 2009/11/15

    Y1 - 2009/11/15

    N2 - The protein kinase Tpl2 (tumour progression locus 2) is activated by LPS (lipopolysaccharide), TNF alpha (tumour necrosis factor alpha) and IL (interleukin)-1. Activation of the native Tpl2 complex by these agonists requires the IKK beta {I kappa B [inhibitor of NF-kappa B (nuclear factor kappa B)] kinase beta}-catalysed phosphorylation of the p105/NF-kappa B1 subunit and is accompanied by the release of the catalytic subunit from both p105/NF-kappa B1 and another subunit ABIN2 (A20-binding inhibitor of NF-kappa B2). In the present study we report that IL-1 activates the transfected Tpl2 catalytic subunit in an HEK (human embryonic kidney)-293 cell line that stably expresses the IL-1R (IL-1 receptor), but does not express the protein kinase IRAK1 (IL-1R-associated kinase). In these cells IL-1 does not activate IKK beta or induce the phosphorylation of p105/NF-kappa B1, and nor does the IKK beta inhibitor PSI 145 prevent the IL-1-induced activation of transfected Tpl2. However, the IL-1-stimulated activation of transfected Tpl2 in IRAK1-null cells or activation of the endogenous Tpl2 complex in IRAK1-expressing cells is suppressed by the protein kinase inhibitor PP2 by a mechanism that does not involve inhibition of Src family protein tyrosine kinases. The IL-1-stimulated activation of transfected Tpl2 is accompanied by its phosphorylation at Thr(290) and Ser(400) and by enhanced phosphorylation of Ser(62), which we demonstrate are autophosphorylation events catalysed by Tpl2 itself. We further show that IL-1 triggers the dissociation of Tpl2 from co-transfected ABIN2 in IRAK1-null IL-1R cells, which is not Suppressed by PP2 or by the inhibition of Tpl2 or IKK beta. These studies identify two new signalling events involved in activation of the native Tpl2 complex by IL-1. First, the IRAK1-, IKK beta- and PP2-independent dissociation of Tpl2 from ABIN2; secondly, the IRAK1- and IKK beta-independent, but PP2-sensitive, activation of the Tpl2 catalytic subunit.

    AB - The protein kinase Tpl2 (tumour progression locus 2) is activated by LPS (lipopolysaccharide), TNF alpha (tumour necrosis factor alpha) and IL (interleukin)-1. Activation of the native Tpl2 complex by these agonists requires the IKK beta {I kappa B [inhibitor of NF-kappa B (nuclear factor kappa B)] kinase beta}-catalysed phosphorylation of the p105/NF-kappa B1 subunit and is accompanied by the release of the catalytic subunit from both p105/NF-kappa B1 and another subunit ABIN2 (A20-binding inhibitor of NF-kappa B2). In the present study we report that IL-1 activates the transfected Tpl2 catalytic subunit in an HEK (human embryonic kidney)-293 cell line that stably expresses the IL-1R (IL-1 receptor), but does not express the protein kinase IRAK1 (IL-1R-associated kinase). In these cells IL-1 does not activate IKK beta or induce the phosphorylation of p105/NF-kappa B1, and nor does the IKK beta inhibitor PSI 145 prevent the IL-1-induced activation of transfected Tpl2. However, the IL-1-stimulated activation of transfected Tpl2 in IRAK1-null cells or activation of the endogenous Tpl2 complex in IRAK1-expressing cells is suppressed by the protein kinase inhibitor PP2 by a mechanism that does not involve inhibition of Src family protein tyrosine kinases. The IL-1-stimulated activation of transfected Tpl2 is accompanied by its phosphorylation at Thr(290) and Ser(400) and by enhanced phosphorylation of Ser(62), which we demonstrate are autophosphorylation events catalysed by Tpl2 itself. We further show that IL-1 triggers the dissociation of Tpl2 from co-transfected ABIN2 in IRAK1-null IL-1R cells, which is not Suppressed by PP2 or by the inhibition of Tpl2 or IKK beta. These studies identify two new signalling events involved in activation of the native Tpl2 complex by IL-1. First, the IRAK1-, IKK beta- and PP2-independent dissociation of Tpl2 from ABIN2; secondly, the IRAK1- and IKK beta-independent, but PP2-sensitive, activation of the Tpl2 catalytic subunit.

    KW - innate immunity

    KW - interleukin-1 (IL-1)

    KW - interleukin-1-receptor-associated kinase 1 (IRAK1)

    KW - nuclear factor kappa B (NF-kappa B)

    KW - tumour necrosis factor (TNF)

    KW - tumour progression locus 2 (Tpl2)

    KW - KAPPA-B-KINASE

    KW - SIGNAL-REGULATED KINASE

    KW - PROTEIN-KINASE

    KW - MAP KINASE

    KW - LIPOPOLYSACCHARIDE ACTIVATION

    KW - NF-KAPPA-B1 P105

    KW - TERNARY COMPLEX

    KW - TNF-ALPHA

    KW - PHOSPHORYLATION

    KW - MACROPHAGES

    U2 - 10.1042/BJ20091271

    DO - 10.1042/BJ20091271

    M3 - Article

    C2 - 19754427

    VL - 424

    SP - 109

    EP - 118

    JO - Biochemical Journal

    JF - Biochemical Journal

    SN - 0264-6021

    ER -