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Identification of duplicated Cited3 genes and their responses to hypoxic stress in blunt snout bream (Megalobrama amblycephala)

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Abstract

The CITED3 protein is a non-DNA-binding transcriptional co-regulator involved in the regulation of various transcriptional responses against hypoxia stress. Here, we characterized two paralogs Cited3 genes (Cited3a and Cited3b) from blunt snout bream (Megalobrama amblycephala), which is a hypoxia-sensitive species. Both genes have an open reading frame of 756 and 723 bp; encoded a protein of 251 amino acid and 240 amino acid, respectively; and they shared a sequence identity of 67%. In adult fish, both Cited3a and Cited3b mRNAs were highly expressed in kidney tissues. In contrast, they were detected in the skin, muscle, and gonad at extraordinarily low levels. During embryogenesis, both Cited3a and Cited3b mRNAs were maternally deposited in eggs and fluctuated from the zygote to the 44-hpf (hours post-fertilization) larvae. Whole-mount in situ hybridization demonstrated that both Cited3a and Cited3b mRNAs were transcribed in the brain, gut, and tailbud at 12 hpf, and at the brain and gut at 24 hpf, and at the brain at 36 hpf embryos. Hypoxic treatment led to upregulated expression of the Cited3 genes during embryogenesis. Under hypoxia, both Cited3a and Cited3b genes in the kidney and brain and Cited3a genes in the liver were significantly upregulated. These results suggest that hypoxia was associated with increases in mRNA levels for both Cited3a (kidney, brain, liver) and Cited3b (kidney and liver).

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References

  • Abraham S, Solomon WB (2000) A novel glutamine-rich putative transcriptional adaptor protein (TIG-1), preferentially expressed in placental and bone-marrow tissues. Gene 255:389–400

    Article  CAS  PubMed  Google Scholar 

  • Bamforth SD, Braganca J, Farthing CR, Schneider JE, Broadbent C, Michell AC, Clarke K, Neubauer S, Norris D, Brown NA, Anderson RH, Bhattacharya S (2004) Cited2 controls left-right patterning and heart development through a Nodal-Pitx2c pathway. Nat Genet 36:1189–1196

    Article  CAS  PubMed  Google Scholar 

  • Bhattacharya S, Ratcliffe PJ (2003) ExCITED about HIF. Nat Struct Biol 10:501–503

    Article  CAS  PubMed  Google Scholar 

  • Bhattacharya S, Michels CL, Leung MK, Arany ZP, Kung AL, Livingston DM (1999) Functional role of p35srj, a novel p300/CBP binding protein, during transactivation by HIF-1. Genes Dev 13:64–75

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bracken CP, Whitelaw ML, Peet DJ (2003) The hypoxia-inducible factors: key transcriptional regulators of hypoxic responses. Cell Mol Life Sci 60:1376–1393

    Article  CAS  PubMed  Google Scholar 

  • Devakanmalai GS, Zumrut HE, Ozbudak EM (2013) Cited3 activates Mef2c to control muscle cell differentiation and survival. Biol Open 2:505–514

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dunwoodie SL, Rodriguez TA, Beddington RS (1998) Msg1 and Mrg1, founding members of a gene family, show distinct patterns of gene expression during mouse embryogenesis. Mech Dev 72:27–40

    Article  CAS  PubMed  Google Scholar 

  • FBMA (2016) Chinese fisheries year book. China Agriculture Press, Beijing

    Google Scholar 

  • Fox SB, Braganca J, Turley H, Campo L, Han C, Gatter KC, Bhattacharya S, Harris AL (2004) CITED4 inhibits hypoxia-activated transcription in cancer cells, and its cytoplasmic location in breast cancer is associated with elevated expression of tumor cell hypoxia-inducible factor 1 alpha. Cancer Res 64:6075–6081

    Article  CAS  PubMed  Google Scholar 

  • Freedman SJ, Sun ZYJ, Kung AL, France DS, Wagner G, Eck MJ (2003) Structural basis for negative regulation of hypoxia-inducible factor-1 alpha by CITED2. Nat Struct Biol 10:504–512

    Article  CAS  PubMed  Google Scholar 

  • Guan WZ, Guo DD, Sun YW, Chen J, Jiang XY, Zou SM (2017) Characterization of duplicated heme oxygenase-1 genes and their responses to hypoxic stress in blunt snout bream (Megalobrama amblycephala). Fish Physiol Biochem 43:641–651

    Article  CAS  PubMed  Google Scholar 

  • Jeon YS, Lee K, Park SC, Kim BS, Cho YJ, Ha SM, Chun J (2014) EzEditor: a versatile sequence alignment editor for both rRNA- and protein-coding genes. Int J Syst Evol Microbiol 64:689–691

    Article  CAS  PubMed  Google Scholar 

  • Jiang XY, Du XD, Tian YM, Shen RJ, Sun CF, Zou SM (2012) Goldfish transposase Tgf2 presumably from recent horizontal transfer is active. FASEB J 26:2743–2752

    Article  CAS  PubMed  Google Scholar 

  • Kinoshita K, Kikuchi Y, Sasakura Y, Suzuki M, Fujii-Kuriyama Y, Sogawa K (2004) Altered DNA binding specificity of Arnt by selection of partner bHLH-PAS proteins. Nucleic Acids Res 32:3169–3179

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Law, S.H.W., Wu, R.S.S., Ng, P.K.S., Yu, R.M.K., Kong, R.Y.C., 2006. Cloning and expression analysis of two distinct HIF-alpha isoforms—gcHIF-1alpha and gcHIF-4alpha—from the hypoxia-tolerant grass carp, Ctenopharyngodon idellus. Bmc Mol Biol 7

  • Li, F.G., Chen, J., Jiang, X.Y., Zou, S.M., 2015. Transcriptome analysis of blunt snout bream (Megalobrama amblycephala) reveals putative differential expression genes related to growth and hypoxia. Plos One 10

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Ng PKS, Wu RSS, Zhang ZP, Mok HOL, Randall DJ, Kong RYC (2003) Molecular cloning and characterization of a hypoxia-responsive CITED3 cDNA from grass carp. Comp Biochem Physiol B 136:163–172

    Article  CAS  PubMed  Google Scholar 

  • Ng, P.K.S., Chiu, S.K., Kwong, T.F.N., Yu, R.M.K., Wong, M.M.L., Kong, R.Y.C., 2009. Functional characterization of two CITED3 homologs (gcCITED3a and gcCITED3b) in the hypoxia-tolerant grass carp, Ctenopharyngodon idellus Bmc Mol Biol 10

  • Ng PKS, Yu RMK, Kwong TFN, Wong MML, Kong RYC (2010) Transcriptional regulation and functional implication of the grass carp CITED1 (gcCITED1) in the negative regulation of HIF-1. Int J Biochem Cell B 42:1544–1552

    Article  CAS  Google Scholar 

  • Semenza GL (2001) HIF-1 and mechanisms of hypoxia sensing. Curr Opin Cell Biol 13:167–171

    Article  CAS  PubMed  Google Scholar 

  • Shen RJ, Jiang XY, Pu JW, Zou SM (2010) HIF-1 alpha and -2 alpha genes in a hypoxia-sensitive teleost species Megalobrama amblycephala: cDNA cloning, expression and different responses to hypoxia. Comp Biochem Physiol B 157:273–280

    Article  CAS  PubMed  Google Scholar 

  • Shi GB, Boyle SC, Sparrow DB, Dunwoodie SL, Shioda T, de Caestecker MP (2006) The transcriptional activity of CITED1 is regulated by phosphorylation in a cell cycle-dependent manner. J Biol Chem 281:27426–27435

    Article  CAS  PubMed  Google Scholar 

  • Sun HB, Zhu YX, Yin T, Sledge G, Yang YC (1998) MRG1, the product of a melanocyte-specific gene related gene, is a cytokine-inducible transcription factor with transformation activity. Proc Natl Acad Sci U S A 95:13555–13560

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sun CF, Tao Y, Jiang XY, Zou SM (2011) IGF binding protein 1 is correlated with hypoxia-induced growth reduce and developmental defects in grass carp (Ctenopharyngodon idellus) embryos. Gen Comp Endocrinol 172:409–415

    Article  CAS  PubMed  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Taylor JS, Braasch I, Frickey T, Meyer A, Van de Peer Y (2003) Genome duplication, a trait shared by 22000 species of ray-finned fish. Genome Res 13:382–390

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Terova G, Rimoldi S, Cora S, Bernardini G, Gornati R, Saroglia M (2008) Acute and chronic hypoxia affects HIF-1 alpha mRNA levels in sea bass (Dicentrarchus labrax). Aquaculture 279:150–159

    Article  CAS  Google Scholar 

  • Thisse, B., 2001. Expression of the zebrafish genome during embryogenesis. Gene expression section (http://zfin.org)

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Weninger WJ, Floro KL, Bennett MB et al (2005) Cited2 is required both for heart morphogenesis and establishment of the left-right axis in mouse development [J]. Development 132(6):1337–1348

    Article  CAS  PubMed  Google Scholar 

  • Wu CB, Liu ZY, Li FG, Chen J, Jiang XY, Zou SM (2017) Gill remodeling in response to hypoxia and temperature occurs in the hypoxia sensitive blunt snout bream (Megalobrama amblycephala). Aquaculture 479:479–486

    Article  CAS  Google Scholar 

  • Yahata T, Shao WL, Endoh H, Hur JY, Coser KR, Sun HP, Ueda Y, Kato S, Isselbacher KJ, Brown M, Shioda T (2001) Selective coactivation of estrogen-dependent transcription by CITED1 CBP/p300-binding protein. Genes Dev 15:2598–2612

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yahata T, Takedatsu H, Dunwoodie SL, Braganca J, Swingler T, Withington SL, Hur J, Coser KR, Isselbacher KJ, Bhattacharya S, Shioda T (2002) Cloning of mouse cited4, a member of the CITED family p300/CBP-binding transcriptional coactivators: induced expression in mammary epithelial cells. Genomics 80:601–613

    Article  CAS  PubMed  Google Scholar 

  • Yin Z, Haynie J, Yang X, Han B, Kiatchoosakun S, Restivo J, Yuan S, Prabhakar NR, Herrup K, Conlon RA, Hoit BD, Watanabe M, Yang YC (2002) The essential role of Cited2, a negative regulator for HIF-1α, in heart development and neurulation. Proc Natl Acad Sci U S A 99(16):10488–10493

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yokota H, Goldring MB, Sun HB (2003) CITED2-mediated regulation of MMP-1 and MMP-13 in human chondrocytes under flow shear. J Biol Chem 278:47275–47280

    Article  CAS  PubMed  Google Scholar 

  • Zhang XL, Sun YW, Chen J, Jiang XY, Zou SM (2017) Gene duplication, conservation and divergence of Heme oxygenase 2 genes in blunt snout bream (Megalobrama amblycephala) and their responses to hypoxia. Gene 610:133–139

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (31572220), the Key Technologies Research and Development Program of China (2012BAD26B02), and the Shanghai University Knowledge Service Platform (ZF1206).

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Correspondence to Xia-Yun Jiang or Shu-Ming Zou.

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Guo, HH., Sun, Y., Zhang, Xl. et al. Identification of duplicated Cited3 genes and their responses to hypoxic stress in blunt snout bream (Megalobrama amblycephala). Fish Physiol Biochem 45, 1141–1152 (2019). https://doi.org/10.1007/s10695-019-00625-6

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  • DOI: https://doi.org/10.1007/s10695-019-00625-6

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