title: Androgen response element of the glycine N-methyltransferase gene is located in the coding region of its first exon.
abstract: Androgen plays an important role in the pathogenesis of PCa (prostate cancer). Previously, we identified GNMT (glycine N-methyltransferase) as a tumour susceptibility gene and characterized its promoter region. Besides, its enzymatic product-sarcosine has been recognized as a marker for prognosis of PCa. The goals of this study were to determine whether GNMT is regulated by androgen and to map its AREs (androgen response elements). Real-time PCR analyses showed that R1881, a synthetic AR (androgen receptor) agonist induced GNMT expression in AR-positive LNCaP cells, but not in AR-negative DU145 cells. In silico prediction showed that there are four putative AREs in GNMT-ARE1, ARE2 and ARE3 are located in the intron 1 and ARE4 is in the intron 2. Consensus ARE motif deduced from published AREs was used to identify the fifth ARE-ARE5 in the coding region of exon 1. Luciferase reporter assay found that only ARE5 mediated the transcriptional activation of R1881. ARE3 overlaps with a YY1 [Yin and Yang 1 (motif (CaCCATGTT, +1118/+1126)] that was further confirmed by antibody supershift and ChIP (chromatin immunoprecipitation) assays. EMSA (electrophoretic mobility shift assay) and ChIP assay confirmed that AR interacts with ARE5 in vitro and in vivo. In summary, GNMT is an AR-targeted gene with its functional ARE located at +19/+33 of the first exon. These results are valuable for the study of the influence of androgen on the gene expression of GNMT especially in the pathogenesis of cancer.
authors.affiliation: *AIDS Prevention and Research Center, National Yang-Ming University, Taipei, Taiwan 11221.
authors.forename: Cheng-Ming
authors.initials: CM
authors.lastname: Lee
authors.forename: Chia-Hung
authors.initials: CH
authors.lastname: Yen
authors.affiliation: §VYM Genome Research Center, National Yang-Ming University, Taipei, Taiwan 11221.
authors.forename: Tsai-Yu
authors.initials: TY
authors.lastname: Tzeng
authors.affiliation: *AIDS Prevention and Research Center, National Yang-Ming University, Taipei, Taiwan 11221.
authors.forename: Yu-Zen
authors.initials: YZ
authors.lastname: Huang
authors.affiliation: §VYM Genome Research Center, National Yang-Ming University, Taipei, Taiwan 11221.
authors.forename: Kuan-Hsien
authors.initials: KH
authors.lastname: Chou
authors.forename: Tai-Jay
authors.initials: TJ
authors.lastname: Chang
authors.forename: Yi-Ming
authors.initials: YM
authors.lastname: Arthur Chen
mesh_terms: D059002:Androgen Receptor Antagonists / Q000494:pharmacology; D000813:Anilides / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D005091:Exons; D015972:Gene Expression Regulation, Neoplastic; D050938:Glycine N-Methyltransferase / Q000235:genetics*; D006801:Humans; D008297:Male; D015741:Metribolone / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D016366:Open Reading Frames; D011471:Prostatic Neoplasms; D011485:Protein Binding; D011944:Receptors, Androgen / Q000378:metabolism; D020218:Response Elements; D017422:Sequence Analysis, DNA; D045165:Testosterone Congeners / Q000494:pharmacology; D014105:Tosyl Compounds / Q000494:pharmacology; D014158:Transcription, Genetic
chemical_list: D059002:Androgen Receptor Antagonists; D000813:Anilides; D009570:Nitriles; D011944:Receptors, Androgen; D045165:Testosterone Congeners; D014105:Tosyl Compounds; D015741:Metribolone; C053541:bicalutamide; C494510:GNMT protein, human; D050938:Glycine N-Methyltransferase