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2013 | 43 | 3 |

Tytuł artykułu

Molecular cloning and expression analysis of hypoxia inducible factor 1α in tongue sole, Cynoglossus semilaevis (Actinopterygii: Pleuronectiformes: Cynoglossidae), subjected to acute hypoxia

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Background. Hypoxia-inducible factor-1α (HIF-1α), a subunit of the HIF-1 protein, plays a key role in the regulation of genes involved in hypoxia physiological response. Tongue sole, Cynoglossus semilaevis Günther, 1873, a marine teleost, had been proved to be a hypoxia-tolerant species. In this study, the HIF-1α in tongue sole was cloned and its expression under acute hypoxia was examined to provide further basis for understanding the molecular response of tongue sole under hypoxia. Materials and Methods. The full length of HIF-1α cDNA sequence was cloned from the liver of tongue sole by RT-PCR and RACE-PCR method. Then, the expression pattern of tongue sole HIF-1α under acute hypoxic conditions were detected using quantitative real-time PCR method. Results. The open reading frame of tongue sole HIF-1α is 2208 bp, encoding 735 amino acids. The amino acid sequence of tongue sole HIF-1α shared high identities (52.7%–81.8%) with HIF-1α from other vertebrates, and possessed six typical domains of the HIF-1 family (bHLH, PAS-A, PAS-B, PAC, N-TAD, and C-TAD). In adult fish, HIF-1α mRNAs were highly expressed in the liver, moderately in the heart, spleen, kidney, stomach, blood and gills, and low in intestine. Under acute hypoxia stress, expression of HIF-1α mRNAs were significantly up-regulated in many tissues, including the liver, spleen, stomach, blood, heart and gills. Conclusion. Tongue sole HIF-1α possessed the similar sequence length, shared higher identities and clustered well with other known HIF-1α, thus revealing a high degree conservation of HIF-1α during evolution. Tongue sole HIF-1α began to be up-regulated from 5 min to 120 min after hypoxia, indicating that it might play a significant role at the early stage of hypoxia.

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  • Post-doctoral research station at Wuxi Fisheries College, Nanjing Agricultural University, 214182, Wuxi, Jiangsu, China
  • Key Laboratory of Aquaculture and Ecology of Coastal pool of Jiangsu Province, Department of Ocean Technology, Yancheng Institute of Technology, Yancheng, Jiangsu 224051, China
  • Key Laboratory of Aquaculture and Ecology of Coastal pool of Jiangsu Province, Department of Ocean Technology, Yancheng Institute of Technology, Yancheng, Jiangsu 224051, China
  • School of Chemical and Biological Engineering, Yancheng Institute of Technology, Yancheng, Jiangsu 224051, China
  • Key Laboratory of Aquaculture and Ecology of Coastal pool of Jiangsu Province, Department of Ocean Technology, Yancheng Institute of Technology, Yancheng, Jiangsu 224051, China
  • Key Laboratory of Aquaculture and Ecology of Coastal pool of Jiangsu Province, Department of Ocean Technology, Yancheng Institute of Technology, Yancheng, Jiangsu 224051, China
  • Key Laboratory of Aquaculture and Ecology of Coastal pool of Jiangsu Province, Department of Ocean Technology, Yancheng Institute of Technology, Yancheng, Jiangsu 224051, China


  • Cao Y.B., Chen X.Q., Wang S., Wang Y. X., Du J.Z. 2008.Evolution and regulation of the downstream gene of hypoxiainducible factor-1α in naked carp (Gymnocypris przewalskii)from lake Qinghai, China. Journal of Molecular Evolution 67 (5): 570–580. DOI: 10.1007/s00239-008-9175-4
  • Chen N., Chen L.P., Zhang J., Chen C., Wei X.L., Gul Y.,Wang W.M., Wang H.L. 2011. Molecular characterization and expression analysis of three hypoxia-inducible factor alpha subunits, HIF-1α/2α/3α of the hypoxia-sensitive freshwater species, Chinese sucker. Gene 498 (1): 81–90. DOI:10.1016/j.gene.2011.12.058
  • Clewley J.P., Arnold C. 1997. Megalign: The multiple alignment module of LASERGENE. Pp. 119–129. In: Swindell S.R. (ed.) Sequence Data Analysis Guidebook. Series: Methods In Molecular Biology. Vol. 70. Springer, New York, USA.DOI: 10.1385/0-89603-358-9:119
  • Gassmann M., Chilov D., Wenger R.H. 2002. Regulation of the hypoxia-inducible factor-1α. ARNT is not necessary for hypoxic induction of HIF-1α in the nucleus. Pp. 87–99. In: Lahiri S.,Prabhakar N.R., Forster R.E.II (eds.) Oxygen sensing: molekule to man. Series: Advances in Experimental Medicine and Biology.Vol. 475. SpringerUS.DOI: 10.1007/0-306-46825-5_9
  • Heidbreder M., Fröhlich F., Jöhren O., Dendorfer A., Oadri F.,Dominiak P. 2003. Hypoxia rapidly activates HIF-3α mRNA expression. The FASEB Journal 17 (11): 1541–1543. DOI: 10.1096/fj.02-0963fje
  • Kageyama Y.,Koshiji M., ToK.K.W., Tian Y.M.,Ratcliffe P.J.,Huang L.E. 2004. Leu-574 of human HIF-1α is a molecular determinant of prolyl hydroxylation. The FASEB Journal 18 (9): 1028–1030. DOI: 10.1096/fj.03-1233fje
  • Kewley R.J., Whitelaw M.L., Chapman-Smith A. 2004. The mammalian basic helix-loop-helix/PAS family of transcriptional regulators. International Journal of Biochemistry and Cell Biology 36 (2): 189–204. DOI: 10.1016/S1357-2725(03)00211-5
  • Kodama K., Rahman M.S., Horiguchi T., Thomas P. 2012.Upregulation of hypoxia-inducible factor (HIF)-1α and HIF-2α mRNA levels in dragonet Callionymus Valenciennes exposed to environmental hypoxia in Tokyo Bay.Marine Pollution Bulletin 64 (7): 1339–1347. DOI:10.1016/j.marpolbul.2012.05.002
  • Kumar S., Dudley J., Nei M., Tamura K. 2008. MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Briefings in Bioinformatics 9 (4): 299–306. DOI: 10.1093/bib/bbn017
  • Law S.H.W., Wu R.S.S., Ng P.K.S., Yu R.M., Kong R.Y. 2006. Cloning and expression analysis of two distinct HIFalpha isoforms—gcHIF-1alpha and gcHIF-4alpha—from the hypoxia-tolerant grass carp, Ctenopharyngodon idellus. BMC Molecular Biology 7 [paper 15]: [13 pages]. DOI: 10.1186/1471-2199-7-15
  • Luo J.C., Shibuya M. 2001. A variant of nuclear localization signal of bipartite-type is required for the nuclear translocation of hypoxia inducible factors (1, 2 and 3). Oncogene 20 (12): 1435–1444.
  • Manolescu B., Oprea E., Busu C., Cercasov C. 2009. Natura compounds and the hypoxia-inducible factor (HIF) signalling pathway. Biochimie 91 (11–12): 1347–1358. DOI: 10.1016/j.biochi.2009.08.005
  • Masson N.,Willam C.,Maxwell P.H., Pugh C.W., Ratcliffe P.J. 2001. Independent function of two destruction domains In hypoxia-inducible factor-α chains activated by propyl hydroxylation. The EMBO Journal 20 (18): 5197–5206.DOI: 10.1093/emboj/20.18.5197
  • Maynard M.A., Ohh M. 2004. von Hippel-Lindau tumor suppressor protein and hypoxia-inducible factor in kidney cancer.American Journal of Nephrology 24 (1): 1–13. DOI:10.1159/000075346
  • Morin P.J., Storey K.B. 2005. Cloning and expression of hypoxia-inducible factor 1 alpha from the hibernating ground squirrel, Sermophilus tridecemlineatus. Biochimica et Biophysica Acta 1729(1): 32–40. DOI: 10.1016/j.bbaexp.2005.02.009
  • Pereira T., Zheng X., Ruas J.L., Tanimoto K., Poellinger L.2003. Identification of residues critical for regulation of protein stability and the transactivation function of the hypoxiainducible factor-1α by the von Hippel-Lindau tumor suppressor gene product. Journal of Biological Chemistry 278 (9): 6816–6823. DOI: 10.1074/jbc.M209297200
  • Qi Z.T., Zhang Q.H., Wang Z.S., Wang A.M., Huang B.,Chang M.X., Nie P. 2011. Cloning and expression analysis of a long type peptidoglycan recognition protein (PGRP-L) from Xenopus tropicalis. Zoological Research 32 (4): 371–378. DOI: 10.3724/SP.J.1141.2011.04371
  • Rahman M.S., Thomas P. 2007. Molecular cloning, characterization and expression of two hypoxia-inducible factor Ralpha subunits, HIF-1α and HIF-2α, in a hypoxia-tolerant marine teleost, Atlantic croaker (Micropogonias undulatus). Gene 396 (2): 273–282. DOI: 10.1016/j.gene.2007.03.009
  • Ratcliffe P.J. 2007. HIF-1 and HIF-2: working alone or together in hypoxia? Journal of Clinical Investigation 117 (4): 862–865. DOI: 10.1172/JCI31750
  • Rojas D.A., Perez-Munizaga D.A., Centanin L., Antonelli M.,Wappner P., Allende M.L., Reyes A.E. 2007. Cloning of hif-1α and hif-2α and mRNA expression pattern during development in zebrafish. Gene Expression Patterns 7 (3): 339–345. DOI: 10.1016/j.modgep.2006.08.002
  • Ruas J.L., Poellinger L., Pereira T. 2002. Functional analysis of hypoxia-inducible factor-1α-mediated transactivation. Identification of amino acid residues critical for transcriptional activation and/or interaction with CREB-binding protein.Journal of Biological Chemistry 277 (41):38723–38730. DOI: 10.1074/jbc.M205051200
  • Rytkönen K.T., Vuori K.A.M., Primmer C.R., Nikinmaa M.2007. Comparison of hypoxia-inducible factor-1 alpha In hypoxia-sensitive and hypoxia-tolerant fish species. Comparative Biochemistry and Physiology Part D:Genomics and Proteomics 2 (2): 177–186. DOI: 10.1016/j.cbd.2007.03.001
  • Shao C.W., Chen S.L., Scheuring C.F., Xu J.Y., Sha Z.X.,Dong X.L., Zhang H.B. 2010. Construction of two BAC libraries from half-smooth tongue sole Cynoglossus semilaevis and identification of clones containing candidate sexdetermination genes. Marine Biotechnology 12 (5): 558–568. DOI: 10.1007/s10126-009-9242-x
  • Shen R.J., Jiang X.Y., Pu J.W., Zou S.M. 2010. HIF-1α and -2α genes in a hypoxia-sensitive teleost species Megalobrama amblycephala: cDNA cloning, expression and different responses to hypoxia. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 157 (3): 273–280. DOI:10.1016/j.cbpb.2010.06.013
  • Soitamo A.J., Rĺbergh C.M.I., Gassmann M., Sistonen L., Nikinmaa M. 2001. Characterization of a hypoxiainducible factor (HIF-1α) from rainbow trout. Accumulation of protein occurs at normal venous oxygen tension. Journal of Biological Chemistry 276 (23): 19699–19705. DOI:10.1074/jbc.M009057200
  • Terova G., Rimoldi S., Corŕ S., Bernardini G., Gornati R.,Saroglia M. 2008. Acute and chronic hypoxia affects HIF-1α mRNA levels in sea bass (Dicentrarchus labrax).Aquaculture 279 (1–4): 150–159. DOI: 10.1016/j.aquaculture.2008.03.041
  • Wang G.L., Jiang B.H., Rue E.A., Semenza G.L. 1995.Hypoxia inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proceedings of the National Academy of Science of the United States of America 92 (12): 5510–5514.
  • Wang Z.S.,QiZ.T.,Qiu M.,Zhao W.H.,Wang A.M.,Huang J.T.,Guo X.J. 2011. Changes of globins expression in tongue sole, Cynoglossus semilaevis (Actinopterygii:Pleuronectiformes: Cynoglossidae) in response to shortterm hypoxia. Acta Ichthyological et Piscatoria 41 (3):179–184. DOI: 10.3750/AIP2011.41.3.05
  • Wang Z.S.,QiZ. T.,Hang Q.H.,Qiu M.,Huang J.T.,Zhao W.H.,Guo X.J. 2012. The cDNA sequence of the transferrin gene in tongue sole (Cynoglossus semilaevis) and its transcriptional changes under acute hypoxic stress. Italian Journal of Animal Science 11 (4): 419–425. DOI: 10.4081/ijas.2012.e74


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