Molecular characterization and tissue expression profile of the Dnmts gene family in pig

doi:10.1016/S2095-3119(16)61512-5

Journal of Integrative Agriculture ›› 2017, Vol. 16 ›› Issue (06): 1367-1374.DOI: 10.1016/S2095-3119(16)61512-5

收稿日期:2016-08-26 出版日期:2017-06-20 发布日期:2017-06-08

Molecular characterization and tissue expression profile of the Dnmts gene family in pig

LUO Zong-gang^{1, 2, 3*}, ZHANG Kai^4*, CHEN Lei^1, ³, YANG Yuan-xin², FU Peng-hui², WANG Ke-tian¹, WANG Ling², LI Ming-zhou⁵, LI Xue-wei⁵, ZUO Fu-yuan², WANG Jin-yong^1, ³

¹Chongqing Academy of Animal Science, Chongqing 402460, P.R.China
²Department of Animal Science, Southwest University (Rongchang Campus), Chongqing 402460, P.R.China
³Key Laboratory of Pig Industry Sciences, Ministry of Agriculture, Chongqing 402460, P.R.China
⁴Sichuan Academy of Grassland Sciences, Chengdu 610000, P.R.China
⁵Institute of Animal Genetics & Breeding, College of Animal Science & Technology, Sichuan Agricultural University, Chengdu 611130, P.R.China

Received:2016-08-26 Online:2017-06-20 Published:2017-06-08
Contact: WANG Jin-yong, Tel: +86-23-46792073, E-mail: dnmtpig@163.com
Supported by:
This work was supported by the grants from the Specialized Doctor Research Fund of Southwestern University of China (2013Bsr8) and the Youth Fund of Rongchang Campus of Southwest University, China (20700429), the Chongqing Fund of Application and Development, China (cstc2013yykfC80003), the Chongqing Fund of Basic Research, China (15428, 14440), and the National Biological Breeding Capacity Building and Industrialization Projects, China (2014-2573).

摘要/Abstract

Abstract: DNA methyltransferases (Dnmts) comprise a family of proteins which involved in the establishment and maintenance of DNA methylation patterns. In pig, the molecular characterization and tissue expression profile of Dnmt gene family are not clear. To solve this problem, reverse transcriptase PCR and rapid amplification of cDNA ends were used to clone the sequences of the porcine Dnmt2 and Dnmt3b genes. Furthermore, the mRNA expression profiles of Dnmt1, Dnmt2, Dnmt3a and Dnmt3b genes from 54 adult tissues and 2 entire fetuses of Rongchang pig were analyzed by quantitative real-time PCR (qRT-PCR). As a result, the lengths of porcine Dnmt2 and Dnmt3b gene cDNAs were 1 227 and 2 559 bp with cytosine-C5 specific DNA methylase domain, respectively. The four Dnmt genes were highly expressed in longissimus dorsi muscle (P<0.01). Dnmt1 is highly expressed in heart (P<0.01) and Dnmt 2 shows its preference in liver and seminal vesicle tissue (P<0.01). Dnmt3a and Dnmt3b are highly expressed in the two fetus stages (P<0.01). All these results suggested that each gene has its specific expression profile, and deeper study is required to dig more details between the methylation level and Dnmt family mRNA expressions in different tissues.

Key words: pig, Dnmt gene family, cloning, mRNA expression

. [J]. Journal of Integrative Agriculture, 2017, 16(06): 1367-1374.

LUO Zong-gang, ZHANG Kai, CHEN Lei, YANG Yuan-xin, FU Peng-hui, WANG Ke-tian, WANG Ling, LI Ming-zhou, LI Xue-wei, ZUO Fu-yuan, WANG Jin-yong. Molecular characterization and tissue expression profile of the Dnmts gene family in pig[J]. Journal of Integrative Agriculture, 2017, 16(06): 1367-1374.

参考文献

Bartolomei M S, Tilghman S M. 1997. Genomic imprinting in mammals. Annual Review of Genetics, 31, 493–525.
Bestor T H. 2000. The DNA methyltransferases of mammals. Human Molecular Genetics, 9, 2395–2402.
Chung Y G, Ratnam S, Chaillet J R, Latham K E. 2003. Abnormal regulation of DNA methyltransferase expression in cloned mouse embryos. Biology of Reproduction, 69, 146–153.
Dong A, Yoder J A, Zhang X, Zhou L, Bestor T H, Cheng X. 2001. Structure of human DNMT2, an enigmatic DNA methyltransferase homolog that displays denaturant-resistant binding to DNA. Nucleic Acids Research, 29, 439–448.
Erkens T, Van Poucke M, Vandesompele J, Goossens K, Van Zeveren A, Peelman L J. 2006. Development of a new set of reference genes for normalization of real-time RT-PCR data of porcine backfat and longissimus dorsi muscle, and evaluation with PPARGC1A. BMC Biotechnology, 6, 41.
Golding M C, Westhusin M E. 2003. Analysis of DNA (cytosine 5)

methyltransferase mRNA sequence and expression in bovine preimplantation embryos, fetal and adult tissues. Gene Expression Patterns, 3, 551–558.
Goll M G, Kirpekar F, Maggert K A, Yoder J A, Hsieh C L, Zhang X, Golic K G, Jacobsen S E, Bestor T H. 2006. Methylation of tRNAAsp by the DNA methyltransferase homolog Dnmt2. Science, 311, 395–398.
Gruenbaum Y, Stein R, Cedar H, Razin A. 1981. Methylation of CpG sequences in eukaryotic DNA. FEBS Letters, 124, 67–71.
Hermann A, Schmitt S, Jeltsch A. 2003. The human Dnmt2 has residual DNA-(cytosine-C5) methyltransferase activity. The Journal of Biological Chemistry, 278, 31717–31721.
Huan Y, Wang H, Wu Z, Zhang J, Liu Z, He H. 2015. The expression patterns of DNA methylation reprogramming related genes are associated with the developmental competence of cloned embryos after zygotic genome activation in pigs. Gene Expression Patterns, 18, 1–7.
Jaenisch R, Bird A. 2003. Epigenetic regulation of gene expression: How the genome integrates intrinsic and environmental signals. Nature Genetics, 33(Suppl.), 245–254.
Jeong Y S, Oh K B, Park J S, Kim J S, Kang Y K. 2009. Cytoplasmic localization of oocyte-specific variant of porcine DNA methyltransferase-1 during early development. Developmental Dynamics, 238, 1666–1673.
Lee J T. 2003. Molecular links between X-inactivation and autosomal imprinting: X-inactivation as a driving force for the evolution of imprinting? Current Biology, 13, R242–R254.
Leonhardt H, Page A W, Weier H U, Bestor T H. 1992. A targeting sequence directs DNA methyltransferase to sites of DNA replication in mammalian nuclei. Cell, 71, 865–873.
Li E, Bestor T H, Jaenisch R. 1992. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell, 69, 915–926.
Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2^–ΔΔCT Method. Methods, 25, 402–408.
Okano M, Bell D W, Haber D A, Li E. 1999. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell, 99, 247–257.
Okano M, Xie S, Li E. 1998a. Cloning and characterization of a family of novel mammalian DNA (cytosine-5) methyltransferases. Nature Genetics, 19, 219–220.
Okano M, Xie S, Li E. 1998b. Dnmt2 is not required for de novo and maintenance methylation of viral DNA in embryonic stem cells. Nucleic Acids Research, 26, 2536–2540.
Prokhortchouk E, Defossez P A. 2008. The cell biology of DNA methylation in mammals. Biochimica et Biophysica Acta, 1783, 2167–2173.
Saradalekshmi K R, Neetha N V, Sathyan S, Nair I V, Nair C M, Banerjee M. 2014. DNA methyl transferase (DNMT) gene polymorphisms could be a primary event in epigenetic susceptibility to schizophrenia. PLoS ONE, 9, e98182.
Schaefer M, Steringer J P, Lyko F. 2008. The Drosophila cytosine-5 methyltransferase Dnmt2 is associated with the nuclear matrix and can access DNA during mitosis. PLoS ONE, 3, e1414.
Sturn A, Quackenbush J, Trajanoski Z. 2002. Genesis: Cluster analysis of microarray data. Bioinformatics, 18, 207–208.
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F. 2002. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology, 3, H34.
Wu H, Zhang Y. 2014. Reversing DNA methylation: Mechanisms, genomics, and biological functions. Cell, 156, 45–68.
Xie S, Wang Z, Okano M, Nogami M, Li Y, He W W, Okumura K, Li E. 1999. Cloning, expression and chromosome locations of the human DNMT3 gene family. Gene, 236, 87–95.
Yoder J A, Bestor T H. 1998. A candidate mammalian DNA methyltransferase related to pmt1p of fission yeast. Human Molecular Genetics, 7, 279–284.
Yoder J A, Soman N S, Verdine G L, Bestor T H. 1997. DNA (cytosine-5)-methyltransferases in mouse cells and tissues. Studies with a mechanism-based probe. Journal of Molecular Biology, 270, 385–395.

Molecular characterization and tissue expression profile of the Dnmts gene family in pig

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