DNA methylation and Yin Yang-1 repress adenosine A₂A receptor levels in human brain

• Published in: Journal of neurochemistry 2010-10, Vol.115 (1), p.283-295
• Main Authors: Buira, Sandra P, Dentesano, Guido, Albasanz, José Luis, Moreno, Jesús, Martín, Mairena, Ferrer, Isidre, Barrachina, Marta
• Format: Article
• Language: English
• Subjects: 5' Untranslated Regions - genetics; Adenosine; Adenosine A2A receptors; Adenylate cyclase; ADORA2A; Aged; Base Sequence; Biological and medical sciences; Blotting, Western; Brain; Brain Chemistry - physiology; Cell Line; Cell receptors; Cell structures and functions; Cerebellum; Cerebellum - metabolism; Chromatin; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; Deoxyribonucleic acid; DNA; DNA Methylation; DNA-Binding Proteins - genetics; Female; Fundamental and applied biological sciences. Psychology; G protein-coupled receptors; g-Aminobutyric acid; Genetic Vectors; Humans; Immunoprecipitation; Male; Medical sciences; Middle Aged; Molecular and cellular biology; Molecular modelling; Molecular Sequence Data; Monoamines receptors (catecholamine, serotonine, histamine, acetylcholine); Neostriatum; Neurochemistry; Neurology; Neurons; Proteins; Putamen; Putamen - metabolism; Receptor, Adenosine A2A - metabolism; Repressors; Reverse Transcriptase Polymerase Chain Reaction; RNA - biosynthesis; RNA - isolation & purification; RNA, Small Interfering - genetics; Transcription factors; Transcription Factors - genetics; Transfection; Western blotting; YY1; YY1 protein; YY1 Transcription Factor - genetics; ZBP-89
• ISSN: 0022-3042; 1471-4159
• DOI: 10.1111/j.1471-4159.2010.06928.x

J. Neurochem. (2010) 115, 283-295. Adenosine A₂A receptors (A₂ARs) are G-protein coupled receptors that stimulate adenylyl cyclase activity. The most A₂ARs-enriched brain region is the striatum, in which A₂ARs are largely restricted to GABAergic neurons of the indirect pathway. We recently described how DNA methylation controls basal A₂AR expression levels in human cell lines. The present report provides clues about the molecular mechanisms that promote human brain region-specific A₂AR gene (ADORA2A) basal expression. The transcription factors ZBP-89 and Yin Yang-1 (YY1) have been characterized as regulators of ADORA2A in SH-SY5Y cells by means of specific expression vectors/siRNAs transient transfection and chromatin immunoprecipitation assay. ZBP-89 plays a role as an activator and YY1 as a repressor. No differences were found in ZBP-89 levels with western blot between the putamen and cerebellum of human postmortem brains. However, increased YY1 levels and DNA methylation percentage in the 5′ untranslated region of ADORA2A, using SEQUENOM MassArray, were found in the cerebellum with respect to the putamen of human brains, showing an inverse relationship with A₂AR levels in the two cerebral regions.