DNA Methylation Variability in a Single Locus of the RXRα Promoter from Umbilical Vein Blood at Term Pregnancy

Diana Chávez-Lizárraga, Paola Zárate-Segura, Jorge Beltrán-Montoya, Carmen Canchola-Sotelo, Felipe Vadillo-Ortega, Erika Chavira-Suárez

Research output: Contribution to journalArticle

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Abstract

© 2018, Springer Science+Business Media, LLC, part of Springer Nature. 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.
Original languageAmerican English
Pages (from-to)210-224
Number of pages187
JournalBiochemical Genetics
DOIs
StatePublished - 1 Jun 2018

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Umbilical Veins
umbilical veins
methylation
DNA methylation
DNA Methylation
pregnancy
Blood
blood
promoter regions
DNA
Pregnancy
loci
protein
growth response
Methylation
Proteins
proteins
Retinoid X Receptors
Growth
Genes

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Chávez-Lizárraga, D., Zárate-Segura, P., Beltrán-Montoya, J., Canchola-Sotelo, C., Vadillo-Ortega, F., & Chavira-Suárez, E. (2018). DNA Methylation Variability in a Single Locus of the RXRα Promoter from Umbilical Vein Blood at Term Pregnancy. Biochemical Genetics, 210-224. https://doi.org/10.1007/s10528-017-9838-1
Chávez-Lizárraga, Diana ; Zárate-Segura, Paola ; Beltrán-Montoya, Jorge ; Canchola-Sotelo, Carmen ; Vadillo-Ortega, Felipe ; Chavira-Suárez, Erika. / DNA Methylation Variability in a Single Locus of the RXRα Promoter from Umbilical Vein Blood at Term Pregnancy. In: Biochemical Genetics. 2018 ; pp. 210-224.
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Chávez-Lizárraga, D, Zárate-Segura, P, Beltrán-Montoya, J, Canchola-Sotelo, C, Vadillo-Ortega, F & Chavira-Suárez, E 2018, 'DNA Methylation Variability in a Single Locus of the RXRα Promoter from Umbilical Vein Blood at Term Pregnancy', Biochemical Genetics, pp. 210-224. https://doi.org/10.1007/s10528-017-9838-1

DNA Methylation Variability in a Single Locus of the RXRα Promoter from Umbilical Vein Blood at Term Pregnancy. / Chávez-Lizárraga, Diana; Zárate-Segura, Paola; Beltrán-Montoya, Jorge; Canchola-Sotelo, Carmen; Vadillo-Ortega, Felipe; Chavira-Suárez, Erika.

In: Biochemical Genetics, 01.06.2018, p. 210-224.

Research output: Contribution to journalArticle

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AU - Chávez-Lizárraga, Diana

AU - Zárate-Segura, Paola

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AU - Chavira-Suárez, Erika

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N2 - © 2018, Springer Science+Business Media, LLC, part of Springer Nature. 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.

AB - © 2018, Springer Science+Business Media, LLC, part of Springer Nature. 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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