RING finger protein AO7 supports NF-κB-mediated transcription by interacting with the transactivation domain of the p65 subunit

被引:30
作者
Asamitsu, K [1 ]
Tetsuka, T [1 ]
Kanazawa, S [1 ]
Okamoto, T [1 ]
机构
[1] Nagoya City Univ, Grad Sch Med Sci, Dept Mol & Cellular Biol, Mizuho Ku, Nagoya, Aichi 4678601, Japan
关键词
D O I
10.1074/jbc.M211831200
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
In this study, a novel interactor of the p65 subunit ( RelA) of NF-kappaB has been explored by performing yeast two-hybrid screen using the transactivation domain (TAD) of p65 located in the C terminus as bait. We have isolated a RING finger motif-containing protein, AO7, previously identified as an interacting protein with a ubiquitin-conjugating enzyme, Ubc5B. We confirmed the protein-protein interaction between p65 and AO7 in vitro and in vivo and found that the C-terminal region of AO7 is responsible for the interaction with p65 TAD. AO7 was predominantly localized in the nucleus and activated the NF-kappaB-dependent gene expression upon stimulation with IL-1beta or TNF or overexpression of NF-kappaB-inducing kinase. We found that both the RING finger and the C-terminal regions of AO7 were necessary for the transcriptional activation. When cotransfected with plasmids expressing Gal4-p65 fusion proteins containing various functional domains of p65, we found that p65 TAD was essential for the transcriptional activation mediated by AO7. Furthermore, the p65-mediated transactivation was suppressed by a ubiquitination-defective AO7 mutant in which the essential Cys residue within the RING finger motif was substituted by Ser. These data suggest that AO7 interacts with the p65 TAD and modulates its transcriptional activity.
引用
收藏
页码:26879 / 26887
页数:9
相关论文
共 40 条
[1]   The NF-kappa B and I kappa B proteins: New discoveries and insights [J].
Baldwin, AS .
ANNUAL REVIEW OF IMMUNOLOGY, 1996, 14 :649-683
[2]   Gene expression - Emerging roles of ubiquitin in transcription regulation [J].
Conaway, RC ;
Brower, CS ;
Conaway, JW .
SCIENCE, 2002, 296 (5571) :1254-1258
[3]   Lung Kruppel-like factor contains an autoinhibitory domain that regulates its transcriptional activation by binding WWP1, an E3 ubiquitin ligase [J].
Conkright, MD ;
Wani, MA ;
Lingrel, JB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (31) :29299-29306
[4]   The PROSITE database, its status in 2002 [J].
Falquet, L ;
Pagni, M ;
Bucher, P ;
Hulo, N ;
Sigrist, CJA ;
Hofmann, K ;
Bairoch, A .
NUCLEIC ACIDS RESEARCH, 2002, 30 (01) :235-238
[5]   Mdm2 is a RING finger-dependent ubiquitin protein ligase for itself and p53 [J].
Fang, SY ;
Jensen, JP ;
Ludwig, RL ;
Vousden, KH ;
Weissman, AM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (12) :8945-8951
[6]   CREB-binding protein p300 are transcriptional coactivators of p65 [J].
Gerritsen, ME ;
Williams, AJ ;
Neish, AS ;
Moore, S ;
Shi, Y ;
Collins, T .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (07) :2927-2932
[7]   Missing pieces in the NF-κB puzzle [J].
Ghosh, S ;
Karin, M .
CELL, 2002, 109 :S81-S96
[8]  
Imhof MO, 1996, MOL CELL BIOL, V16, P2594
[9]   Evidence that de novo protein synthesis is dispensable for anti-apoptotic effects of NF-κB [J].
Kajino, S ;
Suganuma, M ;
Teranishi, F ;
Takahashi, N ;
Tetsuka, T ;
Ohara, H ;
Itoh, M ;
Okamoto, T .
ONCOGENE, 2000, 19 (18) :2233-2239
[10]   Phosphorylation meets ubiquitination:: The control of NF-κB activity [J].
Karin, M ;
Ben-Neriah, Y .
ANNUAL REVIEW OF IMMUNOLOGY, 2000, 18 :621-+