We have identified a new binding partner of the TGFβ (transforming growth factor-β)-activated protein kinase (TAK1), termed TAB3 (TAK1-binding protein-3), which shares 48% amino acid sequence identity with TAB2. Our results indicate that two distinct TAK1 complexes are present in cells. One comprises TAK1 complexed with TAB1 and TAB2, and the other TAK1 complexed with TAB1 and TAB3. Both complexes are activated in response to tumour necrosis factor-α or interleukin-1 in human epithelial KB cells or bacterial lipopolysaccharide in RAW264.7 macrophages, and are subject to feedback control by stress-activated protein kinase 2a (SAPK2a; also called p38α). The electrophoretic mobility of TAB2 and TAB3 decreases in response to these agonists or osmotic shock, and is reversed by treatment with protein phosphatase-1. The decrease in mobility of TAB3 is prevented if the cells are incubated with SB 203580 before stimulation, but treatment with SB 203580 produces forms of TAB2 with a mobility intermediate between that observed for TAB2 in unstimulated and stimulated cells. Similar results were obtained in embryonic fibroblasts from mice deficient in SAPK2a/p38α. Our results indicate that TAB3 is phosphorylated via the SAPK2a/p38α pathway, whereas TAB2 is phosphorylated at two or more sites by both an SAPK2a/p38α-dependent and an SB 203580-independent kinase. The SAPK2a/p38α-mediated phosphorylation of TAB2 and TAB3 may contribute to the SAPK2a/p38α-mediated feedback control of TAK1 activity that also involves the phosphorylation of TAB1. We also show that the agonist-induced activation of TAK1 complexes requires the phosphorylation of the TAK1 catalytic subunit at a serine/threonine residue(s).
- interleukin-1 (IL-1)
- lipopolysaccharide (LPS)
- p38 MAP kinase
- stress-activated protein kinase 2a (SAPK2a)
- tumour necrosis factor (TNF)
The nucleotide sequence data reported for human TAB3 will appear in DDBJ, EMBL, GenBank® and GSDB Nucleotide Sequence Databases under the accession number AY371491.
Abbreviations used: TGFβ, transforming growth factor-β; TAK1, TGFβ-activated protein kinase; TAB1/TAB2/TAB3, TAK1-binding protein-1, -2 and -3 respectively; SAPK2a, stress-activated protein kinase 2a; IL-1, interleukin-1; TNFα, tumour necrosis factor-α; LPS, lipopolysaccharide; JNK, c-Jun N-terminal kinase; NF-κB, nuclear factor κB; GST, glutathione S-transferase; CUE, coupling of ubiquitin conjugation to endoplasmic reticulum degradation; NZF, Npl4 zinc finger; TRAF, TNF-receptor-associated factor.
- The Biochemical Society, London ©2004