title: Sodium arsenite represses the expression of myogenin in C2C12 mouse myoblast cells through histone modifications and altered expression of Ezh2, Glp, and Igf-1.
abstract: Arsenic is a toxicant commonly found in water systems and chronic exposure can result in adverse developmental effects including increased neonatal death, stillbirths, and miscarriages, low birth weight, and altered locomotor activity. Previous studies indicate that 20 nM sodium arsenite exposure to C2C12 mouse myocyte cells delayed myoblast differentiation due to reduced myogenin expression, the transcription factor that differentiates myoblasts into myotubes. In this study, several mechanisms by which arsenic could alter myogenin expression were examined. Exposing differentiating C2C12 cells to 20 nM arsenic increased H3K9 dimethylation (H3K9me2) and H3K9 trimethylation (H3K9me3) by 3-fold near the transcription start site of myogenin, which is indicative of increased repressive marks, and reduced H3K9 acetylation (H3K9Ac) by 0.5-fold, indicative of reduced permissive marks. Protein expression of Glp or Ehmt1, a H3-K9 methyltransferase, was also increased by 1.6-fold in arsenic-exposed cells. In addition to the altered histone remodeling status on the myogenin promoter, protein and mRNA levels of Igf-1, a myogenic growth factor, were significantly repressed by arsenic exposure. Moreover, a 2-fold induction of Ezh2 expression, and an increased recruitment of Ezh2 (3.3-fold) and Dnmt3a (~2-fold) to the myogenin promoter at the transcription start site (-40 to +42), were detected in the arsenic-treated cells. Together, we conclude that the repressed myogenin expression in arsenic-exposed C2C12 cells was likely due to a combination of reduced expression of Igf-1, enhanced nuclear expression and promoter recruitment of Ezh2, and altered histone remodeling status on myogenin promoter (-40 to +42).
authors.affiliation: Environmental Toxicology Graduate Program, Department of Biological Sciences, Clemson University, 132 Long Hall, Clemson, SC 29634, USA.
authors.forename: Gia-Ming
authors.initials: GM
authors.lastname: Hong
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Bain
mesh_terms: D000818:Animals; D018053:Arsenites / Q000633:toxicity*; D002460:Cell Line; D000071221:Enhancer of Zeste Homolog 2 Protein; D005786:Gene Expression Regulation / Q000187:drug effects*; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics; D006657:Histones / Q000187:drug effects* / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000235:genetics; D051379:Mice; D032446:Myoblasts / Q000187:drug effects / Q000378:metabolism; D018008:Myogenin / Q000187:drug effects* / Q000235:genetics; D063151:Polycomb Repressive Complex 2; D011401:Promoter Regions, Genetic; D017670:Sodium Compounds / Q000633:toxicity*
chemical_list: D018053:Arsenites; D006657:Histones; D018008:Myogenin; D017670:Sodium Compounds; C519530:insulin-like growth factor-1, mouse; C017947:sodium arsenite; D007334:Insulin-Like Growth Factor I; D000071221:Enhancer of Zeste Homolog 2 Protein; C498410:Ezh2 protein, mouse; C513479:GLP protein, mouse; D011495:Histone-Lysine N-Methyltransferase; D063151:Polycomb Repressive Complex 2