title: Osteoponin promoter controlled by DNA methylation: aberrant methylation in cloned porcine genome.
abstract: Cloned animals usually exhibited many defects in physical characteristics or aberrant epigenetic reprogramming, especially in some important organ development. Osteoponin (OPN) is an extracellular-matrix protein involved in heart and bone development and diseases. In this study, we investigated the correlation between OPN mRNA and its promoter methylation changes by the 5-aza-dc treatment in fibroblast cell and promoter assay. Aberrant methylation of porcine OPN was frequently found in different tissues of somatic nuclear transferred cloning pigs, and bisulfite sequence data suggested that the OPN promoter region -2615 to -2239 nucleotides (nt) may be a crucial regulation DNA element. In pig ear fibroblast cell culture study, the demethylation of OPN promoter was found in dose-dependent response of 5-aza-dc treatment and followed the OPN mRNA reexpression. In cloned pig study, discrepant expression pattern was identified in several cloned pig tissues, especially in brain, heart, and ear. Promoter assay data revealed that four methylated CpG sites presenting in the -2615 to -2239 nt region cause significant downregulation of OPN promoter activity. These data suggested that methylation in the OPN promoter plays a crucial role in the regulation of OPN expression that we found in cloned pigs genome.
authors.affiliation: Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan.
authors.forename: Chih-Jie
authors.initials: CJ
authors.lastname: Shen
authors.affiliation: Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan ; School of Medicine, College of Medicine, China Medical University, Taichung 40402, Taiwan ; Department of Otolaryngology Head and Neck Surgery, China Medical University, Taichung 40402, Taiwan.
authors.forename: Yung-An
authors.initials: YA
authors.lastname: Tsou
authors.affiliation: Department of Bioresources, Da-Yeh University, Changhwa 515, Taiwan.
authors.forename: Hsiao-Ling
authors.initials: HL
authors.lastname: Chen
authors.affiliation: Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, College of Pharmacy, China Medical University, Taichung 40402, Taiwan.
authors.forename: Hung-Jin
authors.initials: HJ
authors.lastname: Huang
authors.affiliation: Department of Animal Science and Technology, National Taiwan University, Taipei 106, Taiwan.
authors.forename: Shinn-Chih
authors.initials: SC
authors.lastname: Wu
authors.affiliation: Department of Animal Science and Biotechnology, Tung Hai University, Taichung 407, Taiwan.
authors.forename: Winston T K
authors.initials: WT
authors.lastname: Cheng
authors.affiliation: School of Medicine, College of Medicine, China Medical University, Taichung 40402, Taiwan ; Department of Biomedical Informatics, Asia University, Taichung 41354, Taiwan ; Department of Medical Research, Human Genetic Center, China Medical University Hospital, Taichung 40447, Taiwan ; Research Center for Chinese Medicine & Acupuncture, China Medical University, Taichung 40402, Taiwan.
authors.forename: Calvin Yu-Chian
authors.initials: CY
authors.lastname: Chen
authors.affiliation: Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan ; Rong Hsing Research Center for Translational Medicine and the iEGG Center, National Chung Hsing University, Taichung 402, Taiwan.
authors.forename: Chuan-Mu
authors.initials: CM
authors.lastname: Chen
mesh_terms: D000818:Animals; D019976:Cloning, Organism; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D016678:Genome; D053495:Osteopontin / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics*; D013552:Swine
chemical_list: D012333:RNA, Messenger; D053495:Osteopontin
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