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The  Rho x cluster on the mouse X chromosome contains reproduction-related homeobox genes expressed in a sexually dimorphic manner. We report that two members of the  Rhox  cluster,  Rhox6  and  9 , are regulated by de-methylation of histone H3 at lysine 27 by KDM6A, a histone demethylase with female-biased expression. Consistent with other homeobox genes,  Rhox6  and  9  are in bivalent domains prior to embryonic stem cell differentiation and thus poised for activation. In female mouse ES cells, KDM6A is specifically recruited to  Rhox6  and  9  for gene activation, a process inhibited by  Kdm6a  knockdown in a dose-dependent manner. In contrast, KDM6A occupancy at  Rhox6  and  9  is low in male ES cells and knockdown has no effect on expression. In mouse ovary where  Rhox6  and  9  remain highly expressed, KDM6A occupancy strongly correlates with expression. Our study implicates  Kdm6a , a gene that escapes X inactivation, in the regulation of genes important in reproduction, suggesting that KDM6A may play a role in the etiology of developmental and reproduction-related effects of X chromosome anomalies.

Female Bias in Rhox6 and 9 Regulation by the Histone Demethylase KDM6A