Browse > Article

Non-CpG Methylation of Pre-1 Sequence in Pig SCNT Blastocysts  

Ko, Yeoung-Gyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Im, Gi-Sun (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Lee, Hwi-Cheul (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Cho, Sang-Rae (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Choi, Sun-Ho (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Choe, Chang-Yong (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Lee, Poong-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Cho, Chang-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Cho, Jae-Hyeon (College of Veterinary Medicine, Gyeongsang National University)
Yoo, Young-Hee (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Publication Information
Reproductive and Developmental Biology / v.35, no.1, 2011 , pp. 93-97 More about this Journal
Abstract
Previously, we reported that the osmolarity conditions in the satellite region were affected CpG DNA methylation status while Pre-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. This study was conducted to investigate the DNA methylation status of repeat sequences in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaCl or 0.05 M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. The DNA methylation status of the Pre-1 sequences in blastocysts was characterized using a bisulfite-sequencing method. Intriguingly, in the present study, we found the unique DNA methylation at several non-CpG sequences at the Pre-1 sequences in all groups. The non-CpG methylation was hypermethylated in all three groups, including in vivo group (86.90% of PZM-3; 83.87% of NaCl; 84.82% of sucrose; 90.94% of in vivo embryos). To determine whether certain non-CpG methylated sites were preferentially methylated, we also investigated the methylation degree of CpA, CpT and CpC. Excepting in vivo group, preference of methylation was CpT>CpC>CpA in all three groups investigated. These results indicate that DNA methylation of Pre-1 sequences was hypermethylated in CpG as well as non-CpG site, regardless modification of osmolarity in a culture media.
Keywords
NT blastocyst; Methylation; Pre-1; Non-CpG; Osmolarity;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
연도 인용수 순위
1 고응규, 임기순, 박미령, 우제석, 양병철, 황성수, 이휘철, 이풍연, 조창연, 최순호, 유용희 (2010): 삼투압 배양 조건에 따른 돼지 체세포 복제 배반포에서 Repeats 영역의 DNA 메틸화 변화. 한국동물번식학회지 34:181-184.
2 Jones L, Ratcliff F, Baulcombe DC (2001): RNA-directed transcriptional gene silencing in plants can be inherited independently of the RNA trigger and requires Met1 for maintenance. Curr BioI 11:747-757.   DOI   ScienceOn
3 Kremenskoy M, Kremenska Y, Suzuki M, lmai K, Takahashi S, Hashizume K, Yagi S, Shiota K (2006): Epigenetic characterization of the CpG islands of bovine Leptin and POU5F1 genes in cloned bovine fetuses. J Reprod Dev 52:277-285.   DOI   ScienceOn
4 Lindroth AM, Cao X, Jackson JP, Zilberman D, McCallum CM, Henikoff S, Jacobsen SE (2001): Requirement of CHROMOMETHYLASE3 for maintenance of CpXpG methylation. Science 292: 2077-2080.   DOI
5 Lyko F, Ramsahoye BH, Jaenisch R (2000): DNA methylation in Drosophila melanogaster. Nature 408:538-540.   DOI
6 Ohgane J, Wakayama T, Kogo Y, Senda S, Hattori N, Tanaka S, Yanagimachi R, Shiota K (2001): DNA methylation variation in cloned mice. Genesis 30:45-50.   DOI   ScienceOn
7 Clark SJ, Harrison J, Paul CL, Frommer M (1994): High sensitivity mapping of methylated cytosines. Nucleic Acids Res 22:2990-2997.   DOI   ScienceOn
8 Clark SJ, Harrison J, Frommer M (1995): CpNpG methylation in mammalian cells. Nat Genet 10:20-27.   DOI   ScienceOn
9 Dodge JE, Ramsahoye BH, Wo ZG, Okano M, Li E (2002): De novo methylation of MMLV provirus in embryonic stem cells: CpG versus non-CpG methylation. Gene 289: 41-48.   DOI
10 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.   DOI   ScienceOn
11 Hwang IS, Park MR, Moon HJ, Shim JH, Kim DH, Yang BC, Ko YG, Yang BS, Cheong HT, Im GS (2007): Osmolarity at early culture stage affects development and expression of apoptosis related genes (Bax-a and Bcl-xl) in preimplantation porcine NT embryos. Mol Reprod Dev 75:464-471.
12 Kim SH, Kang YK, Koo DB, Kang MJ, Moon SJ, Lee KK, Han YM (2004): Differential DNA methylation reprogramming of various repetitive sequences in mouse preimplantation embryos. Biochemical and Biophysical Research Communications 324:58-63.   DOI   ScienceOn
13 Ko YG, Nishino K, Battori N, Arai Y, Tanaka S, Shiota K (2005): Stage-by-stage change in DNA methylation status of Dnmt1 locus during mouse early development. J BioI Chem 280:9627-9634.
14 Reik W, Dean W, Walter J (2001): Epigenetic reprogramming in mammalian development. Science 293:1089-1093.   DOI   ScienceOn
15 Park MR, Hwang IS, Shim JH, Moon HJ, Kim DH, Ko YK, Seong HH, Im GS (2008): Effect of osmolarity of culture medium on imprinting and apoptotic gene expression in miniature pig nuclear transfer embryos. Reprod Dev BioI 32(3):183-191.
16 Pelissier T, Thalmeir S, Kempe D, Sanger HL, Wassenegger M (1999): Heavy de novo methylation at symmetrical and non-symmetrical sites is a hallmark of RNA-directed DNA methylation. Nucleic Acids Res 27: 1625-1634.   DOI
17 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.   DOI   ScienceOn
18 Shiota K, Kogo Y, Ohgane J, Imamura T, Urano A, Nishino K, Tanaka S, Hattori N (2002): Epigenetic marks by DNA methylation specific to stem, germ and somatic cells in mice. Genes Cells 7:961-969.   DOI   ScienceOn
19 Shi W, Haaf T (2002): Aberrant methylation patterns at the two-cell stage as an indicator of early developmental failure. Mol Reprod Dev 63:329-334.   DOI   ScienceOn
20 Woodcock DM, Lawler CB, Linsenmeyer ME, Doherty JP, Warren WD (1997): Asymmetric methylation in the hypermethylated CpG promoter region of the human L1 retrotransposon. J BioI Chem 272:7810-7816.   DOI
21 Aoki A, Suetake I, Miyagawa J, Fujio T, Chijiwa T, Sasaki H, Tajima S (2001): Enzymatic properties of de novo-type mouse DNA (cytosine-5) methyltransferases. Nucleic Acids Res 29: 3506-3512.   DOI