TNFAIP1 interacts with KCTD10 to promote the degradation of KCTD10 proteins and inhibit the transcriptional activities of NF-κB and AP-1
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The broad-complex, tramtrack, and bric-a-brac/poxvirus and zinc finger domain-containing protein tumor necrosis factor, alpha-induced protein 1 (TNFAIP1) was first identified as a gene whose expression can be induced by the tumor necrosis factor alpha. Some studies showed that TNFAIP1 may function in DNA replication, apoptosis and human diseases. However, the definite functions and the mechanisms of TNFAIP1 are poorly known. In this study, we performed a yeast two-hybrid assay and used TNFAIP1 as the bait to screen human brain cDNA library. Potassium channel tetramerisation domain containing 10 (KCTD10) was identified as TNFAIP1-interacting partner. The KCTD10–TNFAIP1 interaction was then confirmed by the in vitro GST pull-down assays and the in vivo co-immunoprecipitation and colocalization assays. In addition, protein degradation and ubiquitin assays revealed TNFAIP1 overexpression resulted in ubiquitin-mediated degradation of KCTD10 proteins, which was significantly alleviated with the proteasome inhibitor MG132 treatment. Furthermore, transient transfection assays with two reporters showed that TNFAIP1 and KCTD10 inhibited the transcriptional activities of nuclear factor kappa B (NF-κB) and activating protein-1 reporters. Taken together, our results indicated the novel interaction and function between KCTD10 and TNFAIP1 in human PDIP1 family.
KeywordsTNFAIP1 gene KCTD10 gene BTB domain Protein degradation Transcription inhibition
We thank Dr. Yue Xiong (University of North Carolina at Chapel Hill), for providing Cul-3 and ubiquitin constructs. This work was supported in part by the 973 project of Ministry of Science and Technique of China (No. 2009ZX08010-024B), the Program for The National Natural Science Foundation of China (No. 30900827) and the Science & Technology Department of Hunan Province (No. 2011FJ3140, No. 2011TT2008).
- 8.Mencoboni M, Lerza R, Bogliolo G (1992) Tumor necrosis factor: a cytokine with multiple actions. Recent Prog Med 83:15–17Google Scholar
- 11.Xiong X, Liu X, Hu X, Jiang X, Liu Y, Zhang J (2004) Differential expression analysis of B12 gene after mouse partial hepatectomy. J Nat Sci Hunan Norm Univ 27:71–74Google Scholar
- 15.Zhang W, Zhou A, Yang L, Li H, Zhang J (2006) Expression and preparation of antiserum of human KCTD10 and its expression profile in the cell lines. Acta Laser Biol Sinica 15:496–500Google Scholar
- 28.Prajapati B, Singhal M, Yashwant, Sharma GN, Gupta V (2010) Role of NFkB in various immunological & inflammatory disorders. Int J Toxicol Pharmacol Res 2:35–39Google Scholar
- 37.Ortiz-Lazareno PC, Hernandez-Flores G, Dominguez-Rodriguez JR, Lerma-Diaz JM, Jave-Suarez LF, Aguilar-Lemarroy A, Gomez-Contreras PC, Scott-Algara D, Bravo-Cuellar A (2008) MG132 proteasome inhibitor modulates proinflammatory cytokines production and expression of their receptors in U937 cells: involvement of nuclear factor-kappaB and activator protein-1. Immunology 124:534–541PubMedCrossRefGoogle Scholar