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17β-Estradiol Increases Non-CpG Methylation in Exon 1 of the Rainbow Trout (Oncorhynchus mykiss) MyoD Gene

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Abstract

MyoD is an important myogenic transcription factor necessary for the differentiation of myogenic precursor cells (MPC) to form mature myotubes, a process essential for muscle growth. Epigenetic markers such as CpH methylation are known gene regulators that are important for the differentiation process. In this study, we investigated whether DNA methylation is a potential mechanism associated with the ability of 17β-estradiol (E2) to reduce MyoD gene expression and muscle growth in rainbow trout. Rainbow trout received a single intraperitoneal injection of E2 or the injection vehicle (control). Skeletal muscle was collected 24 h post injection and analyzed for DNA methylation within the MyoD gene and the expression of DNA methyltransferases. CpG islands of the MyoD gene were predicted using MethPrimer software, and these regions were PCR amplified and analyzed using bisulfite sequencing. The percent methylation of the targeted CpG did not differ between control and E2-treated fish. However, percent CpH methylation in the MyoD exon 1 region was elevated with E2 treatment. Two of the methylated CpH sites were located in conserved transcription factor binding motifs, estrogen response element (ERE), and Myc binding site. Quantitative real-time PCR analysis revealed a significant increase in expression of DNA methyltransferases, Dnmt3a and Dnmt3b, in E2-treated muscle, suggesting an increased genome methylation. Differential CpH methylation in MyoD gene of control and E2-treated fish suggests an epigenetic mechanism through which E2 decreases MyoD gene expression and contributes to reduced muscle growth.

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References

  • Akagi T, Henry IM, Kawai T, Comai L, Tao R (2016) Epigenetic regulation of the sex determination gene MeGI in polyploid persimmon. Plant Cell 28:2905–2915

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arand J, Spieler D, Karius T, Branco MR, Meilinger D, Meissner A, Jenuwein T, Xu G, Leonhardt H, Wolf V, Walter J (2012) In vivo control of CpG and non-CpG DNA methylation by DNA methyltransferases. PLoS Genet 8:e1002750

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Avery OT, Macleod CM, Mccarty M (1944) Studies on the chemical nature of the substance inducing transformation of pneumococcal types: induction of transformation by a desoxyribonucleic acid fraction isolated from pneumococcus type III. J Exp Med 79:137–158

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brunk BP, Goldhamer DJ, Emerson JCP (1996) Regulated demethylation of the myoD distal enhancer during skeletal myogenesis. Dev Biol 177:490–503

    Article  CAS  PubMed  Google Scholar 

  • Cheng X, Blumenthal RM (2010) Coordinated chromatin control: structural and functional linkage of DNA and histone methylation. Biochemistry 49:2999–3008

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cleveland BM, Weber GM (2016) Effects of steroid treatment on growth, nutrient partitioning, and expression of genes related to growth and nutrient metabolism in adult triploid rainbow trout (Oncorhynchus mykiss). Domest Anim Endocrinol 56:1–12

    Article  CAS  PubMed  Google Scholar 

  • Compere SJ, Palmiter RD (1981) DNA methylation controls the inducibility of the mouse metallothionein-I gene in lymphoid cells. Cell 25:233–240

    Article  CAS  PubMed  Google Scholar 

  • Fuso A, Ferraguti G, Grandoni F, Ruggeri R, Scarpa S, Strom R, Lucarelli M (2010) Early demethylation of non-CpG, CpC-rich, elements in the myogenin 5′-flanking region. Cell Cycle 9:3965–3976

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gómez S, Castellano G, Mayol G, Suñol M, Queiros A, Bibikova M, Nazor KL, Loring JF, Lemos I, Rodríguez E, De Torres C, Mora J, Martín-Subero JI, Lavarino C (2015) DNA methylation fingerprint of neuroblastoma reveals new biological and clinical insights. Epigenomics 7:1137–1153

    Article  PubMed  Google Scholar 

  • Haines TR, Rodenhiser DI, Ainsworth PJ (2001) Allele-specific non-CpG methylation of the Nf1 gene during early mouse development. Dev Biol 240:585–598

    Article  CAS  PubMed  Google Scholar 

  • Holliday R, Pugh JE (1975) DNA modification mechanisms and gene activity during development. Science 187:226–232

    Article  CAS  PubMed  Google Scholar 

  • Jones PA, Wolkowicz MJ, Harrington MA, Gonzales F (1990) Methylation and expression of the Myo D1 determination gene. Philos Trans R Soc Lond Ser B Biol Sci 326:277

    Article  CAS  Google Scholar 

  • Koganti PP, Wang J, Cleveland B, Ma H, Weber GM, Yao J (2017) Estradiol regulates expression of miRNAs associated with myogenesis in rainbow trout. Mol Cell Endocrinol 443:1–14

    Article  CAS  PubMed  Google Scholar 

  • Kouidou S, Agidou T, Kyrkou A, Andreou A, Katopodi T, Georgiou E, Krikelis D, Dimitriadou A, Spanos P, Tsilikas C, Destouni H, Tzimagiorgis G (2005) Non-CpG cytosine methylation of p53 exon 5 in non-small cell lung carcinoma. Lung Cancer 50:299–307

    Article  PubMed  Google Scholar 

  • Lister R, Mukamel EA, Nery JR, Urich M, Puddifoot CA, Johnson ND, Lucero J, Huang Y, Dwork AJ, Schultz MD, Yu M, Tonti-Filippini J, Heyn H, Hu S, Wu JC, Rao A, Esteller M, He C, Haghighi FG, Sejnowski TJ, Behrens MM, Ecker JR (2013) Global epigenomic reconfiguration during mammalian brain development. Science 341:1237905

  • Manohar A, Anwar RA (1994) Evidence for a cell-specific negative regulatory element in the first intron of the gene for bovine elastin. Biochem J 300:147

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Masser DR, Stanford DR, Freeman WM (2015) Targeted DNA methylation analysis by next-generation sequencing. J Vis Exp 96:e52488

  • Mccarty M, Avery OT (1946) Studies on the chemical nature of the substance inducing transformation of pneumococcal types: ii. Effect of desoxyribonuclease on the biological activity of the transforming substance. J Exp med 83:89–96

    Article  CAS  PubMed Central  Google Scholar 

  • Nazar DS, Persson G, Olin T, Waters S, Von Der Decken A (1991) Sarcoplasmic and myofibrillar proteins in white truck muscle of salmon (Salmo salar) after estradiol treatment. Comp Biochem Physiol B 98B:109–114

    Article  CAS  Google Scholar 

  • Norbeck LA, Sheridan MA (2011) An in vitro model for evaluating peripheral regulation of growth in fish: effects of 17beta-estradiol and testosterone on the expression of growth hormone receptors, insulin-like growth factors, and insulin-like growth factor type 1 receptors in rainbow trout (Oncorhynchus mykiss). Gen Comp Endocrinol 173:270–280

    Article  CAS  PubMed  Google Scholar 

  • Pierce RA, Moore CH, Arikan MC (2006) Positive transcriptional regulatory element located within exon 1 of elastin gene. Am J Physiol Lung Cell Mol Physiol 291:L391

    Article  CAS  PubMed  Google Scholar 

  • Prendergast GC, Lawe D, Ziff EB (1991) Association of Myn, the murine homolog of Max, with c-Myc stimulates methylation-sensitive DNA binding and ras cotransformation. Cell 65:395–407

    Article  CAS  PubMed  Google Scholar 

  • Ramsahoye BH, Biniszkiewicz D, Lyko F, Clark V, Bird AP, Jaenisch R (2000) Non-CpG methylation is prevalent in embryonic stem cells and may be mediated by DNA methyltransferase 3a. Proc Natl Acad Sci 97:5237–5242

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Robinson JT, Thorvaldsdottir H, Winckler W, Guttman M, Lander ES, Getz G, Mesirov JP (2011) Integrative genomics viewer. Nat Biotechnol 29:24–26

  • Schultz MD, He Y, Whitaker JW, Hariharan M, Mukamel EA, Leung D, Rajagopal N, Nery JR, Urich MA, Chen H, Lin S, Lin Y, Jung I, Schmitt AD, Selvaraj S, Ren B, Sejnowski TJ, Wang W, Ecker JR (2015) Human body epigenome maps reveal noncanonical DNA methylation variation. Nature 523:212–216

  • Takagi H, Tajima S, Asano A (1995) Overexpression of DNA methyltransferase in myoblast cells accelerates myotube formation. Eur J Biochem 231:282–291

    Article  CAS  PubMed  Google Scholar 

  • Tomizawa S-I, Kobayashi H, Watanabe T, Andrews S, Hata K, Kelsey G, Sasaki H (2011) Dynamic stage-specific changes in imprinted differentially methylated regions during early mammalian development and prevalence of non-CpG methylation in oocytes. Development 138:811

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wallace RA (1985) Vitellogenesis and oocyte growth in nonmammalian vertebrates. In: Browder LW (ed) Oogenesis. Springer US, Boston

    Google Scholar 

  • Ziller MJ, Müller F, Liao J, Zhang Y, Gu H, Bock C, Boyle P, Epstein CB, Bernstein BE, Lengauer T, Gnirke A, Meissner A (2011) Genomic distribution and inter-sample variation of non-CpG methylation across human cell types. PLoS Genet 7:e1002389

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This study was supported by the USDA ARS Cooperative Agreement No. 58-1930-0-059.

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Authors and Affiliations

  1. Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV, 26506-6108, USA

    Prasanthi P. Koganti & Jianbo Yao

  2. Department of Biological Sciences, University of Delaware, Newark, DE, USA

    Jian Wang

  3. National Center for Cool and Cold Water Aquaculture, USDA/ARS, Kearneysville, WV, USA

    Beth Cleveland

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  1. Prasanthi P. Koganti
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  2. Jian Wang
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  3. Beth Cleveland
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  4. Jianbo Yao
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Correspondence to Jianbo Yao.

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Koganti, P.P., Wang, J., Cleveland, B. et al. 17β-Estradiol Increases Non-CpG Methylation in Exon 1 of the Rainbow Trout (Oncorhynchus mykiss) MyoD Gene. Mar Biotechnol 19, 321–327 (2017). https://doi.org/10.1007/s10126-017-9756-6

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  • DOI: https://doi.org/10.1007/s10126-017-9756-6

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