demethylation of the coding region triggers the activation of the human testis-specific pdha2 gene in somatic tissues脱甲基的编码区触发激活人类testis-specific pdha2基因体细胞组织.pdfVIP

Demethylation of the Coding Region Triggers the Activation of the Human Testis-Specific PDHA2 Gene in Somatic Tissues 1 ˜ 1 ˆ 1 1,2 ˜ 1,2 Ana Pinheiro , Maria Joao Nunes , Ines Milagre , Elsa Rodrigues , Maria Joao Silva , Isabel Tavares de Almeida1,2, Isabel Rivera1,2* 1 Research Institute for Medicines and Pharmaceutical Sciences (iMed.UL), Lisbon, Portugal, 2 Department of Biochemistry and Human Biology, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal Abstract Human PDHA2 is a testis-specific gene that codes for the E a subunit of Pyruvate Dehydrogenase Complex (PDC), a crucial 1 enzyme system in cell energy metabolism. Since activation of the PDHA2 gene in somatic cells could be a new therapeutic approach for PDC deficiency, we aimed to identify the regulatory mechanisms underlying the human PDHA2 gene expression. Functional deletion studies revealed that the 2122 to 26 promoter region is indispensable for basal expression of this TATA-less promoter, and suggested a role of an epigenetic program in the control of PDHA2 gene expression. Indeed, treatment of SH-SY5Y cells with the hypomethylating agent 5-Aza-2 9-deoxycytidine (DAC) promoted the reactivation of the PDHA2 gene, by inducing the recruitment of the RNA polymerase II to the proximal promoter region and the consequent increase in PDHA2 mRNA levels. Bisulfite sequencing analysis revealed that DAC treatment induced a significant demethylation of the CpG island II (nucleotides + 197 to +460) in PDHA2 coding region, while the promoter region remained highly methylated. Taken togethe