Intermolecular Binding Between TIFA-FHA and TIFA-pT Mediates Tumor Necrosis Factor Alpha Stimulation and NF-κB Activation

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2012 May 9

PMID 22566686

Citations 23

Authors

Chia-Chi Flora Huang

Jui-Hung Weng

Tong-You Wade Wei

Pei-Yu Gabriel Wu

Pang-Hung Hsu

Yu-Hou Chen

Shun-Chang Wang

Dongyan Qin

Chin-Chun Hung

Shui-Tsung Chen

Andrew H-J Wang

John Y-J Shyy

Ming-Daw Tsai

Affiliations

Soon will be listed here.

Abstract

The forkhead-associated (FHA) domain recognizes phosphothreonine (pT) with high specificity and functional diversity. TIFA (TRAF-interacting protein with an FHA domain) is the smallest FHA-containing human protein. Its overexpression was previously suggested to provoke NF-κB activation, yet its exact roles in this signaling pathway and the underlying molecular mechanism remain unclear. Here we identify a novel threonine phosphorylation site on TIFA and show that this phosphorylated threonine (pT) binds with the FHA domain of TIFA, leading to TIFA oligomerization and TIFA-mediated NF-κB activation. Detailed analysis indicated that unphosphorylated TIFA exists as an intrinsic dimer and that the FHA-pT9 binding occurs between different dimers of TIFA. In addition, silencing of endogenous TIFA resulted in attenuation of tumor necrosis factor alpha (TNF-α)-mediated downstream signaling. We therefore propose that the TIFA FHA-pT9 binding provides a previously unidentified link between TNF-α stimulation and NF-κB activation. The intermolecular FHA-pT9 binding between dimers also represents a new mechanism for the FHA domain.

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References

Sun Z, Hsiao J, Fay D, Stern D . Rad53 FHA domain associated with phosphorylated Rad9 in the DNA damage checkpoint. Science. 1998; 281(5374):272-4. DOI: 10.1126/science.281.5374.272. View

Zapata J, Pawlowski K, Haas E, Ware C, Godzik A, Reed J . A diverse family of proteins containing tumor necrosis factor receptor-associated factor domains. J Biol Chem. 2001; 276(26):24242-52. DOI: 10.1074/jbc.M100354200. View

Liang X, Van Doren S . Mechanistic insights into phosphoprotein-binding FHA domains. Acc Chem Res. 2008; 41(8):991-9. PMC: 2962622. DOI: 10.1021/ar700148u. View

Wang K, Galson D, Auron P . TRAF6 is autoinhibited by an intramolecular interaction which is counteracted by trans-ubiquitination. J Cell Biochem. 2010; 110(3):763-71. PMC: 3407548. DOI: 10.1002/jcb.22589. View

Baud V, Liu Z, Bennett B, Suzuki N, Xia Y, Karin M . Signaling by proinflammatory cytokines: oligomerization of TRAF2 and TRAF6 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain. Genes Dev. 1999; 13(10):1297-308. PMC: 316725. DOI: 10.1101/gad.13.10.1297. View

Fan A, Deb-Basu D, Orban M, Gotlib J, Natkunam Y, ONeill R . Nanofluidic proteomic assay for serial analysis of oncoprotein activation in clinical specimens. Nat Med. 2009; 15(5):566-71. PMC: 4006986. DOI: 10.1038/nm.1903. View

Force W, Glass A, Benedict C, Cheung T, Lama J, Ware C . Discrete signaling regions in the lymphotoxin-beta receptor for tumor necrosis factor receptor-associated factor binding, subcellular localization, and activation of cell death and NF-kappaB pathways. J Biol Chem. 2001; 275(15):11121-9. DOI: 10.1074/jbc.275.15.11121. View

Fang C, Chen H, Wang M, Chen P, Chang C, Chen L . Global analysis of modifications of the human BK virus structural proteins by LC-MS/MS. Virology. 2010; 402(1):164-76. DOI: 10.1016/j.virol.2010.03.029. View

Durocher D, Jackson S . The FHA domain. FEBS Lett. 2002; 513(1):58-66. DOI: 10.1016/s0014-5793(01)03294-x. View

10.

Inoue J, Yagi S, Ishikawa K, Azuma S, Ikawa S, Semba K . Identification and characterization of Xenopus laevis homologs of mammalian TRAF6 and its binding protein TIFA. Gene. 2005; 358:53-9. DOI: 10.1016/j.gene.2005.05.016. View

11.

Kanamori M, Suzuki H, Saito R, Muramatsu M, Hayashizaki Y . T2BP, a novel TRAF2 binding protein, can activate NF-kappaB and AP-1 without TNF stimulation. Biochem Biophys Res Commun. 2002; 290(3):1108-13. DOI: 10.1006/bbrc.2001.6315. View

12.

Xu X, Tsvetkov L, Stern D . Chk2 activation and phosphorylation-dependent oligomerization. Mol Cell Biol. 2002; 22(12):4419-32. PMC: 133858. DOI: 10.1128/MCB.22.12.4419-4432.2002. View

13.

Li J, Taylor I, Lloyd J, Clapperton J, Howell S, Macmillan D . Chk2 oligomerization studied by phosphopeptide ligation: implications for regulation and phosphodependent interactions. J Biol Chem. 2008; 283(51):36019-30. DOI: 10.1074/jbc.M804075200. View

14.

Barthe P, Roumestand C, Canova M, Kremer L, Hurard C, Molle V . Dynamic and structural characterization of a bacterial FHA protein reveals a new autoinhibition mechanism. Structure. 2009; 17(4):568-78. DOI: 10.1016/j.str.2009.02.012. View

15.

ONeill R, Bhamidipati A, Bi X, Deb-Basu D, Cahill L, Ferrante J . Isoelectric focusing technology quantifies protein signaling in 25 cells. Proc Natl Acad Sci U S A. 2006; 103(44):16153-8. PMC: 1618307. DOI: 10.1073/pnas.0607973103. View

16.

Ishida T, MIZUSHIMA S, Azuma S, Kobayashi N, Tojo T, Suzuki K . Identification of TRAF6, a novel tumor necrosis factor receptor-associated factor protein that mediates signaling from an amino-terminal domain of the CD40 cytoplasmic region. J Biol Chem. 1996; 271(46):28745-8. DOI: 10.1074/jbc.271.46.28745. View

17.

Liu J, Luo S, Zhao H, Liao J, Li J, Yang C . Structural mechanism of the phosphorylation-dependent dimerization of the MDC1 forkhead-associated domain. Nucleic Acids Res. 2012; 40(9):3898-912. PMC: 3351156. DOI: 10.1093/nar/gkr1296. View

18.

Takatsuna H, Kato H, Gohda J, Akiyama T, Moriya A, Okamoto Y . Identification of TIFA as an adapter protein that links tumor necrosis factor receptor-associated factor 6 (TRAF6) to interleukin-1 (IL-1) receptor-associated kinase-1 (IRAK-1) in IL-1 receptor signaling. J Biol Chem. 2003; 278(14):12144-50. DOI: 10.1074/jbc.M300720200. View

19.

Jungmichel S, Clapperton J, Lloyd J, Hari F, Spycher C, Pavic L . The molecular basis of ATM-dependent dimerization of the Mdc1 DNA damage checkpoint mediator. Nucleic Acids Res. 2012; 40(9):3913-28. PMC: 3351161. DOI: 10.1093/nar/gkr1300. View

20.

Pullen S, Miller H, Everdeen D, Dang T, Crute J, Kehry M . CD40-tumor necrosis factor receptor-associated factor (TRAF) interactions: regulation of CD40 signaling through multiple TRAF binding sites and TRAF hetero-oligomerization. Biochemistry. 1998; 37(34):11836-45. DOI: 10.1021/bi981067q. View