Abstract
DNA methylation status of RXRα gene promoter has been correlated with maternal diet during early pregnancy, and associated with offspring’s adiposity and bone mineral content. In adult life, increased methylation of RXRα promoter region is associated with myocardium pathologies. Early growth response proteins (EGR) are zinc finger transcription factors associated with several cellular pathways such as inflammation, apoptosis, and cardiopathies. DNA-binding sequences of EGR proteins have been reported in the RXRα gene promoter using chromatin immunoprecipitation methods. Here, we used correlations between the maternal pre-pregnancy body mass index (p-BMI), gestational weight gain (GWG), and birth weight (BW) as indirect indicators of the maternal nutritional status as modifier of DNA methylation in the offspring. DNA methylation status from newborns’ umbilical vein blood in full-term pregnancy was evaluated in a short sequence (116 pb) of the RXRα gene promoter that contains the elements of response sequence for EGR proteins. Fifty-three bisulfite-modified DNA samples were assessed through methyl-sensitive high-resolution melting (MS-HRM) analysis. To validate the results, we directly sequenced MS-HRM samples to confirm the presence of CpG-methylated positions. In addition, the RXRα protein levels in extracts of umbilical vein blood were evaluated by western blot. We found differential methylation in a specific locus of the RXRα promoter surrounding the EGR-binding sequence; however, no correlation was found with the level of RXRα protein expression. Variability in the methylation status of the RXRα promoter near the EGR transcription factor binding site in newborn cord blood provides controversial epigenetic insights into RXRα regulation via EGR proteins.
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Acknowledgements
This project was supported by the Dirección General de Asuntos del Personal Académico (DGAPA), UNAM postdoctoral fellowship for ECS, and funds from Fundación Miguel Alemán, A.C. We would like to thank Soledad López-Alvarez and Beatriz Sánchez-Hernández (CINVESTAV-IPN and INCMNSZ) for technical assistance.
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The authors declare that there are no known conflicts of interest associated with this publication. The present project was conducted with full compliance of research ethics norms, and more specifically the rules, specifications, and good practices established in the Official Mexican Standards for health and by the Mexico City Ministry of Health.
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Chávez-Lizárraga, D., Zárate-Segura, P., Beltrán-Montoya, J. et al. DNA Methylation Variability in a Single Locus of the RXRα Promoter from Umbilical Vein Blood at Term Pregnancy. Biochem Genet 56, 210–224 (2018). https://doi.org/10.1007/s10528-017-9838-1
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DOI: https://doi.org/10.1007/s10528-017-9838-1