RRC ID 47436
Author Nakaoka HJ, Hara T, Yoshino S, Kanamori A, Matsui Y, Shimamura T, Sato H, Murakami Y, Seiki M, Sakamoto T.
Title NECAB3 Promotes Activation of Hypoxia-inducible factor-1 during Normoxia and Enhances Tumourigenicity of Cancer Cells.
Journal Sci Rep
Abstract Unlike most cells, cancer cells activate hypoxia inducible factor-1 (HIF-1) to use glycolysis even at normal oxygen levels, or normoxia. Therefore, HIF-1 is an attractive target in cancer therapy. However, the regulation of HIF-1 during normoxia is not well characterised, although Mint3 was recently found to activate HIF-1 in cancer cells and macrophages by suppressing the HIF-1 inhibitor, factor inhibiting HIF-1 (FIH-1). In this study, we analysed Mint3-binding proteins to investigate the mechanism by which Mint3 regulates HIF-1. Yeast two-hybrid screening using Mint3 as bait identified N-terminal EF-hand calcium binding protein 3 (NECAB3) as a novel factor regulating HIF-1 activity via Mint3. NECAB3 bound to the phosphotyrosine-binding domain of Mint3, formed a ternary complex with Mint3 and FIH-1, and co-localised with Mint3 at the Golgi apparatus. Depletion of NECAB3 decreased the expression of HIF-1 target genes and reduced glycolysis in normoxic cancer cells. NECAB3 mutants that binds Mint3 but lacks an intact monooxygenase domain also inhibited HIF-1 activation. Inhibition of NECAB3 in cancer cells by either expressing shRNAs or generating a dominant negative mutant reduced tumourigenicity. Taken together, the data indicate that NECAB3 is a promising new target for cancer therapy.
Volume 6
Pages 22784
Published 2016-3-7
DOI 10.1038/srep22784
PII srep22784
PMID 26948053
PMC PMC4780031
MeSH Adaptor Proteins, Signal Transducing Binding Sites Carcinogenesis / genetics Carcinogenesis / metabolism* Carrier Proteins / chemistry Carrier Proteins / genetics Carrier Proteins / metabolism* Cell Line, Tumor Glycolysis Golgi Apparatus / metabolism HEK293 Cells Humans Hypoxia-Inducible Factor 1 / metabolism* Mixed Function Oxygenases / metabolism* Repressor Proteins / metabolism* Ternary Complex Factors
IF 3.998
Times Cited 7
DNA material pcDNA3.1_V5-NECAB3_WT (RDB14306) pcDNA3.1_V5-NECAB3_N1 (RDB14307) pcDNA3.1_V5-NECAB3_N2 (RDB14308) pcDNA3.1_V5-NECAB3_C1 (RDB14309) pcDNA3.1_V5-NECAB3_C2 (RDB14310) pcDNA3.1_V5-NECAB3_C3 (RDB14311) pcDNA3.1_V5-NECAB3_H/A (RDB14312) pcDNA3.1_V5-Mint3_WT-FLAG (RDB14313) pcDNA3.1_V5-Mint3_N-FLAG (RDB14314) pcDNA3.1_V5-Mint3_C-FLAG (RDB14315) pcDNA3.1_V5-Mint3_dPDZ1-FLAG (RDB14316) pcDNA3.1_V5-Mint3_dPDZ2-FLAG (RDB14317) pcDNA3.1_V5-Mint3_dPTB-FLAG (RDB14318) pcDNA3.1_V5-Mint3_PTB-FLAG (RDB14319) pcDNA3.2_MTI-MMP_Myc (RDB14320) pcDNA3.2_Mint3_Myc (RDB14321) pcDNA3.2_FIH-1_FLAG (RDB14322) pLenti6_FLAG-NECAB3 (RDB14305).