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Molecular response and association analysis of Megalobrama amblycephala fih-1 with hypoxia

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Abstract

Hypoxia is one of the most important environmental factors which affect fish growth, development and survival, but regulation mechanisms of hypoxia in fish remain unclear. Therefore, to further understand molecular functions of factor inhibiting HIF-1 (Fih-1), an essential hypoxia sensor, the full-length cDNA of fih-1 was cloned from Megalobrama amblycephala, a hypoxia-sensitive cyprinid fish. The deduced amino acid sequence showed high homology with that of other vertebrates, and all structural and functional domains were highly conserved. The mRNA level in different tissues and developmental stages indicated that M. amblycephala fih-1 expression was higher in liver and muscle, followed by gill, intestine and spleen. During embryogenesis, the fih-1 mRNA was highly expressed in the early embryonic development, then decreased to a very low level, and maintained a relative high level of expression after hatching. In most tissues, the fih-1 mRNA was down-regulated at 2 h but up-regulated at 4 h after hypoxia treatment. In addition, the promoter sequence of M. amblycephala fih-1 was obtained using thermal asymmetric interlaced PCR. Three single nucleotide polymorphism (SNP) sites were found in the cDNA and promoter sequences, and identified significant association with hypoxia trait by correlation analysis in hypoxia-sensitive group and hypoxia-tolerant group. These results demonstrated that M. amblycephala fih-1 plays important roles in embryo development and hypoxia response, which will contribute to systematic understanding of the molecular mechanisms of fish in response to hypoxia, and provide help for fish genetic breeding with hypoxia-tolerant strains or breeds.

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References

  • Barrick D, Kopan R (2006) The Notch transcription activation complex makes its move. Cell 124:883–885

    Article  CAS  PubMed  Google Scholar 

  • Belozeroy VE, Van Meir EG (2005) Hypoxia inducible factor-1: a novel target for cancer therapy. Anticancer Drugs 16:901–909

    Article  Google Scholar 

  • Bray S (1998) A Notch affair. Cell 93:499–503

    Article  CAS  PubMed  Google Scholar 

  • Buckler ES, Thornsberry JM (2002) Plant molecular diversity and applications to genomics. Curr Opin Plant Biol 5:107–111

    Article  CAS  PubMed  Google Scholar 

  • Chen N, Huang CH, Chen BX, Liu H, Wang WM, Gul Y, Wang HL (2015) Alternative splicing transcription of Megalobrama amblycephala HIF prolyl hydroxylase PHD3 and up-regulation of PHD3 by HIF-1alpha. Biochem Biophys Res Commun 469:737–742

    Article  PubMed  Google Scholar 

  • Cockman ME, Webb JD, Kramer HB, Kessler BM, Ratcliffe PJ (2009) Proteomics-based identification of novel factor inhibiting hypoxia-inducible factor (FIH) substrates indicates widespread asparaginyl hydroxylation of ankyrin repeat domain-containing proteins. Mol Cell Proteomics 8:535–546

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Coleman ML, McDonough MA, Hewitson KS, Coles C, Mecinovic J, Edelmann M, Cook KM, Cockman ME, Lancaster DE, Kessler BM, Oldham NJ, Ratcliffe PJ, Schofield CJ (2007) Asparaginyl hydroxylation of the Notch ankyrin repeat domain by factor inhibiting hypoxia-inducible factor. J Biol Chem 282:24027–24038

    Article  CAS  PubMed  Google Scholar 

  • Dayan F, Roux D, Brahimi-Horn MC, Pouyssegur J, Mazure NM (2006) The oxygen sensor factor-inhibiting hypoxia-inducible factor-1 controls expression of distinct genes through the bifunctional transcriptional character of hypoxia-inducible factor-1alpha. Cancer Res 66:3688–3698

    Article  CAS  PubMed  Google Scholar 

  • Dayan F, Monticelli M, Pouysségur J, Pécou E (2009) Gene regulation in response to graded hypoxia: the non-redundant roles of the oxygen sensors and FIH in the HIF pathway. J Theor Biol 259:304–316

    Article  CAS  PubMed  Google Scholar 

  • Ding Z, Wu J, Su L, Zhou F, Zhao X, Deng W, Zhang J, Liu S, Wang W, Liu H (2013) Expression of heat shock protein 90 genes during early development and infection in Megalobrama amblycephala and evidence for adaptive evolution in teleost. Dev Comp Immunol 41:683–693

    Article  CAS  PubMed  Google Scholar 

  • Elson DA, Thurston G, Huang LE, Ginzinger DG, McDonald DM, Johnson RS, Arbeit JM (2001) Induction of hypervascularity without leakage or inflammation in transgenic mice overexpressing hypoxia-inducible factor-1alpha. Genes Dev 15:2520–2532

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Freedman SJ, Sun ZY, Poy F, Kung AL, Livingston DM, Wagner G, Eck MJ (2002) Structural basis for recruitment of CBP/p300 by hypoxia-inducible factor-1 alpha. Proc Natl Acad Sci USA 99:5367–5372

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Geng X, Feng J, Liu S, Wang Y, Arias C, Liu Z (2014) Transcriptional regulation of hypoxia inducible factors alpha (HIF-alpha) and their inhibiting factor (FIH-1) of channel catfish (Ictalurus punctatus) under hypoxia. Comp Biochem Physiol B Biochem Mol Biol 169:38–50

    Article  CAS  PubMed  Google Scholar 

  • Goto Y, Yue L, Yokoi A, Nishimura R, Uehara T, Koizumi S, Saikawa Y (2001) A novel single-nucleotide polymorphism in the 3′-untranslated region of the human dihydrofolate reductase gene with enhanced expression. Clin Cancer Res 7:1952–1956

    CAS  PubMed  Google Scholar 

  • Guo SW, Thompson EA (1992) Performing the exact test of Hardy–Weinberg proportion for multiple alleles. Biometrics 48:361–372

    Article  CAS  PubMed  Google Scholar 

  • Hewitson KS, McNeill LA, Riordan MV, Tian YM, Bullock AN, Welford RW, Elkins JM, Oldham NJ, Bhattacharya S, Gleadle JM, Ratcliffe PJ, Pugh CW, Schofield CJ (2002) Hypoxia-inducible factor (HIF) asparagine hydroxylase is identical to factor inhibiting HIF (FIH) and is related to the cupin structural family. J Biol Chem 277:26351–26355

    Article  CAS  PubMed  Google Scholar 

  • Hirota K, Semenza GL (2005) Regulation of hypoxia-inducible factor 1 by prolyl and asparaginyl hydroxylases. Biochem Biophys Res Commun 338:610–616

    Article  CAS  PubMed  Google Scholar 

  • Huang LE, Bunn HF (2003) Hypoxia-inducible factor and its biomedical relevance. J Biol Chem 278:19575–19578

    Article  CAS  PubMed  Google Scholar 

  • Huang LE, Gu J, Schau M, Bunn HF (1998) Regulation of hypoxia-inducible factor 1a is mediated by an O2-dependent degradation domain via the ubiquitin–proteasome pathway. Proc Natl Acad Sci USA 95:7987–7992

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hyseni A, van der Groep P, van der Wall E, van Diest PJ (2011) Subcellular FIH-1 expression patterns in invasive breast cancer in relation to HIF-1alpha expression. Cell Oncol (Dordr) 34:565–570

    Article  CAS  Google Scholar 

  • Koivunen P, Hirsila M, Gunzler V, Kivirikko KI, Myllyharju J (2004) Catalytic properties of the asparaginyl hydroxylase (FIH) in the oxygen sensing pathway are distinct from those of its prolyl 4-hydroxylases. J Biol Chem 279:9899–9904

    Article  CAS  PubMed  Google Scholar 

  • Kuzmanov A, Wielockx B, Rezaei M, Kettelhake A, Breier G (2012) Overexpression of factor inhibiting HIF-1 enhances vessel maturation and tumor growth via platelet-derived growth factor-C. Int J Cancer 131:E603–E613

    Article  CAS  PubMed  Google Scholar 

  • Lando D, Peet DJ, Gorman JJ, Whelan DA, Whitelaw ML, Bruick R (2002a) FIH-1 is a an asparaginyl hydroxylase that regulates the transcriptional activity of hypoxia inducible factor. Genes Dev 16:1466–1471

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lando D, Peet DJ, Whelan DA, Gorman JJ, Whitelaw ML (2002b) Asparagine hydroxylation of the HIF transactivation domain a hypoxic switch. Science 295:858–861

    Article  CAS  PubMed  Google Scholar 

  • Layman CA, Smith DE, Herod JD (2000) Seasonally varying importance of abiotic and biotic factors in marsh-pond fish communities. Mar Ecol Prog Ser 207:155–169

    Article  Google Scholar 

  • Lee C, Kim SJ, Jeong DG, Lee SM, Ryu SE (2003) Structure of human FIH-1 reveals a unique active site pocket and interaction sites for HIF-1 and von Hippel-Lindau. J Biol Chem 278:7558–7563

    Article  CAS  PubMed  Google Scholar 

  • Liu YG, Whittier RF (1995) Thermal asymmetric interlaced PCR: automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking. Genomics 25:674–681

    Article  CAS  PubMed  Google Scholar 

  • Liu YG, Mitsukawa N, Oosumi T, Whittier RF (1995) Efficient isolation and mapping of T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J 8:457–463

    Article  CAS  PubMed  Google Scholar 

  • Mahon PC, Hirota K, Semenza GL (2001) FIH-1: a novel protein that interacts with HIF-1alpha and VHL to mediate repression of HIF-1 transcriptional activity. Genes Dev 15:2675–2686

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mumm JS, Kopan R (2000) Notch signaling: from the outside in. Dev Biol 228:151–165

    Article  CAS  PubMed  Google Scholar 

  • Peet DJ, Lando D, Whelan DA, Whitelaw ML, Gorman JJ (2004) Oxygen-dependent asparagine hydroxylation. Methods Enzymol 381:467–487

    Article  CAS  PubMed  Google Scholar 

  • Pollock MS, Clarke LMJ, Dubé MG (2007) The effects of hypoxia on fishes: from ecological relevance to physiological effects. Environ Rev 15:1–14

    Article  CAS  Google Scholar 

  • Resnik ER, Herron JM, Lyu SC, Cornfield DN (2007) Developmental regulation of hypoxia-inducible factor 1 and prolyl-hydroxylases in pulmonary vascular smooth muscle cells. Proc Natl Acad Sci USA 104:18789–18794

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rodriguez S, Gaunt TR, Day IN (2009) Hardy-Weinberg equilibrium testing of biological ascertainment for Mendelian randomization studies. Am J Epidemiol 169:505–514

    Article  PubMed  PubMed Central  Google Scholar 

  • Sainson RC, Harris AL (2006) Hypoxia-regulated differentiation: let’s step it up a Notch. Trends Mol Med 12:141–143

    Article  CAS  PubMed  Google Scholar 

  • Satake H, Sasaki N (2010) Comparative overview of toll-like receptors in lower animals. Zoolog Sci 27:154–161

    Article  CAS  PubMed  Google Scholar 

  • Semenza GL (1999) Perspectives on oxygen sensing. Cell 98:281–284

    Article  CAS  PubMed  Google Scholar 

  • Semenza GL (2004) Hydroxylation of HIF-1: oxygen sensing at the molecular level. Physiology 19:176–182

    Article  CAS  PubMed  Google Scholar 

  • Semenza GL, Agani F, Iyer N, Kotch L, Laughner E, Leung S, Yu A (1999) Regulation of cardiovascular development and physiology by hypoxia-inducible factor 1a. Ann N Y Acad Sci 874:262–268

    Article  CAS  PubMed  Google Scholar 

  • So JH, Kim JD, Yoo KW, Kim HT, Jung SH, Choi JH, Lee MS, Jin SW, Kim CH (2014) FIH-1, a novel interactor of mindbomb, functions as an essential anti-angiogenic factor during zebrafish vascular development. PLoS One 9:e109517

    Article  PubMed  PubMed Central  Google Scholar 

  • Sun T, Gao Y, Tan W, Ma S, Shi Y, Yao J, Guo Y, Yang M, Zhang X, Zhang Q, Zeng C, Lin D (2007) A six-nucleotide insertion-deletion polymorphism in the CASP8 promoter is associated with susceptibility to multiple cancers. Nat Genet 39:605–613

    Article  CAS  PubMed  Google Scholar 

  • Wang E, Zhang C, Polavaram N, Liu F, Wu G, Schroeder MA, Lau JS, Mukhopadhyay D, Jiang SW, O’Neill BP, Datta K, Li J (2014) The role of factor inhibiting HIF (FIH-1) in inhibiting HIF-1 transcriptional activity in glioblastoma multiforme. PLoS One 9:e86102

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang H, Huang C, Chen N, Zhu K, Chen B, Wang W, Wang H (2015) Molecular characterization and mRNA expression of HIF-prolyl hydroxylase-2 (phd2) in hypoxia-sensing pathways from Megalobrama amblycephala. Comp Biochem Physiol B Biochem Mol Biol 186:28–35

    Article  CAS  PubMed  Google Scholar 

  • Welker TL, Mcnulty ST, Klesius PH (2007) Effect of sublethal hypoxia on the immune response and susceptibility of channel catfish, Ictalurus punctatus, to enteric septicemia. J World Aquacult Soc 38:12–23

    Article  Google Scholar 

  • Wilkins SE, Hyvarinen J, Chicher J, Gorman JJ, Peet DJ, Bilton RL, Koivunen P (2009) Differences in hydroxylation and binding of Notch and HIF-1alpha demonstrate substrate selectivity for factor inhibiting HIF-1 (FIH-1). Int J Biochem Cell Biol 41:1563–1571

    Article  CAS  PubMed  Google Scholar 

  • Xiao W (2015) The hypoxia signaling pathway and hypoxic adaptation in fishes. Sci China Life Sci 58:148–155

    Article  CAS  PubMed  Google Scholar 

  • Zhang N, Fu Z, Linke S, Chicher J, Gorman JJ, Visk D, Haddad GG, Poellinger L, Peet DJ, Powell F, Johnson RS (2010) The asparaginyl hydroxylase factor inhibiting HIF-1alpha is an essential regulator of metabolism. Cell Metab 11:364–378

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zheng X, Linke S, Dias JM, Zheng X, Gradin K, Wallis TP, Hamilton BR, Gustafsson M, Ruas JL, Wilkins S, Bilton RL, Brismar K, Whitelaw ML, Pereira T, Gorman JJ, Ericson J, Peet DJ, Lendahl U, Poellinger L (2008) Interaction with factor inhibiting HIF-1 defines an additional mode of cross-coupling between the Notch and hypoxia signaling pathways. Proc Natl Acad Sci USA 105:3368–3373

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

This research was supported by the Fundamental Research Funds for the Central Universities (2013PY067 and 2662015PY205), Key Technologies R&D Program of China (no. 2012BAD26B00), and Modern Agroindustry Technology Research System entitled “Staple Freshwater Fishery Industry Technology System” (no. CARS-46-05).

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Authors and Affiliations

  1. Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Bao Zhang, Nan Chen, Cuihong Huang, Chunxiao Huang, Boxiang Chen, Hong Liu, Weimin Wang & Huanling Wang

  2. Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, 430070, People’s Republic of China

    Boxiang Chen, Hong Liu & Huanling Wang

  3. Department of Zoology, Government College Women University Faisalabad, Faisalabad, Pakistan

    Yasmeen Gul

  4. Hubei BaiRong Improved Aquatic Seed Co., Ltd, Huanggang, 438800, People’s Republic of China

    Boxiang Chen

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  1. Bao Zhang
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Correspondence to Huanling Wang.

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All the experimental procedures were approved by the institution of animal care and use committee of Huazhong Agricultural University.

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Communicated by J. Cerdá.

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Zhang, B., Chen, N., Huang, C. et al. Molecular response and association analysis of Megalobrama amblycephala fih-1 with hypoxia. Mol Genet Genomics 291, 1615–1624 (2016). https://doi.org/10.1007/s00438-016-1208-x

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  • DOI: https://doi.org/10.1007/s00438-016-1208-x

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