The role of TBK1 and IKKε in the expression and activation of Pellino 1

TY - JOUR

T1 - The role of TBK1 and IKKε in the expression and activation of Pellino 1

AU - Smith, Hilary

AU - Liu, Xin-Yu

AU - Da, Liang

AU - Goh, Eddy T. H.

AU - Chan, Aye-Thu

AU - Xi, Jiajia

AU - Seh, Cheah-Chen

AU - Qureshi, Insaf A.

AU - Lescar, Julien

AU - Ruedl, Christiane

AU - Gourlay, Robert

AU - Morton, Simon

AU - Hough, Joanne

AU - McIver, Edward G.

AU - Cohen, Philip

AU - Cheung, Peter C. F.

PY - 2011/3/15

Y1 - 2011/3/15

N2 - Mammalian Pellino isoforms are phosphorylated by IRAK (interleukin receptor associated kinase) 1/IRAK4 in vitro, converting them into active E3 ubiquitin ligases. In the present paper we report a striking enhancement in both transcription of the gene encoding Pellino I and Pellino 1 protein expression when murine BMDMs (bone-marrow-derived macrophages) are stimulated with LPS (lipopolysaccharide) or poly(l:C). This induction occurs via a TRIF [TIR (Toll/interleukin-1 receptor)-domain-containing adaptor-inducing interferon-beta]-dependent IRAK-independent pathway and is prevented by inhibition of the IKK [I kappa B (inhibitor of nuclear factor kappa B) kinase]related protein kinases, TBKI {TANK [TRAF (tumour-necrosis-factor-receptor-associated factor)-associated nuclear factor kappa B activator]-binding kinase 1} and IKK epsilon. Pellino I is not induced in IRF3 (interferon regulatory factor 3)(-/-) BMDMs, and its induction is only reduced slightly in type 1 interferon receptor(-/-) BMDMs, identifying Pellino 1 as a new IRF3-dependent gene. We also identify Pellino 1 in a two-hybrid screen using 1KK epsilon as bait, and show that IKK epsilon/TBK1 activate Pellino 1 in vitro by phosphorylating Ser(76), Thr(288) and Ser(293). Moreover, we show that the E3 ligase activity of endogenous Pellino 1 is activated in LPS- or poly(I:C)-stimulated macrophages. This occurs more rapidly than the increase in Pellino 1 mRNA and protein expression, is prevented by the inhibition of IKK epsilon/TBKI and is reversed by phosphatase treatment. Thus IKK epsilon/TBK1 mediate the activation of Pellino 1's E3 ligase activity, as well as inducing the transcription of its gene and protein expression in response to TLR3 and TLR4 agonists.

AB - Mammalian Pellino isoforms are phosphorylated by IRAK (interleukin receptor associated kinase) 1/IRAK4 in vitro, converting them into active E3 ubiquitin ligases. In the present paper we report a striking enhancement in both transcription of the gene encoding Pellino I and Pellino 1 protein expression when murine BMDMs (bone-marrow-derived macrophages) are stimulated with LPS (lipopolysaccharide) or poly(l:C). This induction occurs via a TRIF [TIR (Toll/interleukin-1 receptor)-domain-containing adaptor-inducing interferon-beta]-dependent IRAK-independent pathway and is prevented by inhibition of the IKK [I kappa B (inhibitor of nuclear factor kappa B) kinase]related protein kinases, TBKI {TANK [TRAF (tumour-necrosis-factor-receptor-associated factor)-associated nuclear factor kappa B activator]-binding kinase 1} and IKK epsilon. Pellino I is not induced in IRF3 (interferon regulatory factor 3)(-/-) BMDMs, and its induction is only reduced slightly in type 1 interferon receptor(-/-) BMDMs, identifying Pellino 1 as a new IRF3-dependent gene. We also identify Pellino 1 in a two-hybrid screen using 1KK epsilon as bait, and show that IKK epsilon/TBK1 activate Pellino 1 in vitro by phosphorylating Ser(76), Thr(288) and Ser(293). Moreover, we show that the E3 ligase activity of endogenous Pellino 1 is activated in LPS- or poly(I:C)-stimulated macrophages. This occurs more rapidly than the increase in Pellino 1 mRNA and protein expression, is prevented by the inhibition of IKK epsilon/TBKI and is reversed by phosphatase treatment. Thus IKK epsilon/TBK1 mediate the activation of Pellino 1's E3 ligase activity, as well as inducing the transcription of its gene and protein expression in response to TLR3 and TLR4 agonists.

KW - E3 ligase

KW - interferon regulatory factor 3 (IRF3)

KW - interleukin receptor-associated kinase (IRAK)

KW - Toll-like receptor (TLR)

KW - Toll/IL-1 receptor-domain-containing adaptor-inducing interferon-beta (TRIF)

KW - ubiquitin

KW - KAPPA-B ACTIVATION

KW - E3 UBIQUITIN LIGASE

KW - INTERLEUKIN-1 RECEPTOR

KW - LYS63-LINKED POLYUBIQUITINATION

KW - CRYSTAL-STRUCTURES

KW - SIGNALING PATHWAY

KW - MAMMALIAN-CELLS

KW - VIRAL-INFECTION

KW - INNATE IMMUNITY

KW - KINASE

U2 - 10.1042/BJ20101421

DO - 10.1042/BJ20101421

M3 - Article

C2 - 21204785

SN - 0264-6021

VL - 434

SP - 537

EP - 548

JO - Biochemical Journal

JF - Biochemical Journal

ER -