Molecular basis for the unique deubiquitinating activity of the NF-κB inhibitor A20

被引:136
作者
Lin, Su-Chang [1 ]
Chung, Jee Y. [1 ]
Lamothe, Betty [2 ]
Rajashankar, Kanagalaghatta [3 ]
Lu, Miao [1 ]
Lo, Yu-Chih [1 ]
Lam, Amy Y. [1 ]
Darnay, Bryant G. [2 ]
Wu, Hao [1 ]
机构
[1] Cornell Univ, Weill Med Coll, Dept Biochem, New York, NY 10021 USA
[2] Univ Texas Houston, Dept Expt Therapeut, MD Anderson Canc Ctr, Houston, TX 77030 USA
[3] Argonne Natl Lab, NE CAT, Argonne NAtl Lab, Argonne, IL 60439 USA
基金
美国国家卫生研究院;
关键词
A20; crystal structure; deubiquitination; DUB; TRAF6;
D O I
10.1016/j.jmb.2007.11.092
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nuclear factor kappa B (NF-kappa B) activation in tumor necrosis factor, interleukin-1, and Toll-like receptor pathways requires Lys63-linked nondegradative polyubiquitination. A20 is a specific feedback inhibitor of NF-kappa B activation in these pathways that possesses dual ubiquitin-editing functions. While the N-terminal domain of A20 is a deubiquitinating enzyme (DUB) for Lys63-linked polyubiquitinated signaling mediators such as TRAF6 and RIP, its C-terminal domain is a ubiquitin ligase (E3) for Lys48-linked degradative polyubiquitination of the same substrates. To elucidate the molecular basis for the DUB activity of A20, we determined its crystal structure and performed a series of biochemical and cell biological studies. The structure reveals the potential catalytic mechanism of A20, which may be significantly different from papain-like cysteine proteases. Ubiquitin can be docked onto a conserved A20 surface; this interaction exhibits charge complementarity and no steric clash. Surprisingly, A20 does not have specificity for Lys63-linked polyubiquitin chains. Instead, it effectively removes Lys63-linked polyubiquitin chains from TRAF6 without dissembling the chains themselves. Our studies suggest that A20 does not act as a general DUB but has the specificity for particular polyubiquitinated substrates to assure its fidelity in regulating NF-kappa B activation in the tumor necrosis factor, interleukin-1, and Toll-like receptor pathways. (C) 2007 Elsevier Ltd. All rights reserved.
引用
收藏
页码:526 / 540
页数:15
相关论文
共 66 条
[1]   Mechanism and function of deubiquitinating enzymes [J].
Amerik, AY ;
Hochstrasser, M .
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2004, 1695 (1-3) :189-207
[2]   TAK1-dependent signaling requires functional interaction with TAB2/TAB3 [J].
Besse, Arnaud ;
Lamothe, Betty ;
Campos, Alejandro D. ;
Webster, William K. ;
Maddineni, Upendra ;
Lin, Su-Chang ;
Wu, Hao ;
Darnay, Bryant G. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2007, 282 (06) :3918-3928
[3]   A20 and A20-binding proteins as cellular inhibitors of nuclear factor-κB-dependent gene expression and apoptosis [J].
Beyaert, R ;
Heyninck, K ;
Van Huffel, S .
BIOCHEMICAL PHARMACOLOGY, 2000, 60 (08) :1143-1151
[4]   The ubiquitin-modifying enzyme A20 is required for termination of Toll-like receptor responses [J].
Boone, DL ;
Turer, EE ;
Lee, EG ;
Ahmad, RC ;
Wheeler, MT ;
Tsui, C ;
Hurley, P ;
Chien, M ;
Chai, S ;
Hitotsumatsu, O ;
McNally, E ;
Pickart, C ;
Ma, A .
NATURE IMMUNOLOGY, 2004, 5 (10) :1052-1060
[5]   Chemistry-based functional proteomics reveals novel members of the deubiquitinating enzyme [J].
Borodovsky, A ;
Ovaa, H ;
Kolli, N ;
Gan-Erdene, T ;
Wilkinson, KD ;
Ploegh, HL ;
Kessler, BM .
CHEMISTRY & BIOLOGY, 2002, 9 (10) :1149-1159
[6]   Ubiquitin signalling in the NF-κB pathway [J].
Chen, ZJJ .
NATURE CELL BIOLOGY, 2005, 7 (08) :758-U19
[7]   TAB3, a new binding partner of the protein kinase TAK1 [J].
Cheung, PCF ;
Nebreda, AR ;
Cohen, P .
BIOCHEMICAL JOURNAL, 2004, 378 :27-34
[8]   A20 blocks endothelial cell activation through a NF-kappa B-dependent mechanism [J].
Cooper, JT ;
Stroka, DM ;
Brostjan, C ;
Palmetshofer, A ;
Bach, FH ;
Ferran, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (30) :18068-18073
[9]   Kinetic and mechanistic studies on the hydrolysis of ubiquitin C-terminal 7-amido-4-methylcoumarin by deubiquitinating enzymes [J].
Dang, LC ;
Melandri, FD ;
Stein, RL .
BIOCHEMISTRY, 1998, 37 (07) :1868-1879
[10]   Activation of the IκB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain [J].
Deng, L ;
Wang, C ;
Spencer, E ;
Yang, LY ;
Braun, A ;
You, JX ;
Slaughter, C ;
Pickart, C ;
Chen, ZJ .
CELL, 2000, 103 (02) :351-361