title: Epigenetic modifications of gene promoter DNA in the liver of adult female mice masculinized by testosterone.
abstract: Testosterone (T) is known to masculinize the female phenotype of the liver, evidenced as up- and down-regulated expressions of male- and female-predominant genes, respectively, involved in hepatic metabolism. This study is aimed at identifying epigenetic modifications of promoters of these differently expressed genes in the liver after masculinization by T of adult female C57BL/6 mice using methylated DNA immunoprecipitation and NimbleGen microarrays. Among the 17,354 promoters examined, 82 promoters in the liver have been identified to be significantly changed by T (p<0.05), with 47 and 35 promoters exhibiting increased and decreased DNA methylation, respectively. Most of these promoters display the changes of DNA methylation in their Ups-regions, which are between +500 and +2000 bp upstream from the transcription start site (TSS) of the genes. Less T-induced modifications have been detected in the Cor-regions of the promoters, i.e., +500 to -500 bp around the TSS. Only 13 and 7 Cor-promoters are hyper- and hypo-methylated, respectively, among which are 10 hyper- and 5 hypo-methylated promoters of genes with annotated functions. Surprisingly, the promoters are largely unmethylated in those genes whose expression has been previously found to be permanently deregulated by T in the liver, as e.g. the T-upregulated male-predominant genes Cyp7b1, Cyp2d9, Cyp4a10, Ugt2b1, Ugt2b38, Hsd3b5, Slco1a1 as well as the T-downregulated female-predominant genes Cyp2b9, Cyp2b13, Cyp3a41, Cyp3a44, Fmo3, Sult2a2, respectively. Though methylatable, the promoter DNA of Ar, Esr1, and Esr2 remained unaffected by T. However, T decreases DNA-methylation of the Cor-promoter region of Ddc encoding the AR-coactivator dopa decarboxylase. Among the identified 15 Cor-promoters of genes with annotated functions are also those of Defb43, Cst11, and Sele involved in innate immunity. Our data support the view that T may exert long-lasting epigenetic effects on functions of the liver-inherent immune system.
authors.affiliation: Department of Zoology, College of Science, King Saud University, 11451 Riyadh, Saudi Arabia; Department of Zoology and Entomology, Faculty of Science, Helwan University, Cairo, Egypt. Electronic address: mohameddkhil@yahoo.com.
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Dkhil
authors.affiliation: Department of Zoology, College of Science, King Saud University, 11451 Riyadh, Saudi Arabia.
authors.forename: Saleh
authors.initials: S
authors.lastname: Al-Quraishy
authors.affiliation: Department of Zoology, College of Science, King Saud University, 11451 Riyadh, Saudi Arabia; Department of Zoology, Faculty of Science, Beni-Suef University, Beni-Suef, Egypt.
authors.forename: Abdel-Azeem
authors.initials: AA
authors.lastname: Abdel-Baki
authors.affiliation: Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University, Duesseldorf, Germany.
authors.forename: Foued
authors.initials: F
authors.lastname: Ghanjati
authors.affiliation: Computational Biology and Systems Biomedicine, Biodonostia Health Research Institute, San Sebastion, Spain; IKERBASQUE, Basque Foundation for Science, Bilbao, Spain.
authors.forename: Marcos J
authors.initials: MJ
authors.lastname: Arauzo-Bravo
authors.affiliation: Department of Biology, Heinrich-Heine-University, Duesseldorf, Germany; Boehringer-Ingelheim, Biberach, Germany.
authors.forename: Denis
authors.initials: D
authors.lastname: Delic
authors.affiliation: Department of Biology, Heinrich-Heine-University, Duesseldorf, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Wunderlich
mesh_terms: D000818:Animals; D018899:CpG Islands; D019175:DNA Methylation / Q000187:drug effects*; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005260:Female; D047468:Immunoprecipitation; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D011401:Promoter Regions, Genetic; D013739:Testosterone / Q000494:pharmacology*
chemical_list: D013739:Testosterone
keywords: DNA methylomics; Epigenetics; Gene promoter; Liver; Testosterone