DNA methylation regulates adenosine A(2A) receptor cell surface expression levels.
Adenosine A(2A) receptors (A(2A)Rs) appear to play important roles in inflammation and in certain diseases of the nervous system. Pharmacological modulation of A(2A)Rs is particularly useful in Parkinson's disease and has been tested in schizophrenia. However, little is known about the regulation of A(2A)R gene (ADORA2A). A bioinformatic analysis revealed the presence of three CpG islands in the 5' UTR region of human ADORA2A. Next, HeLa, SH-SY5Y and U87-MG cells were treated for 48 h with 5 muM 5-azacytidine (Aza). Increased A(2A)R levels were demonstrated in HeLa and SH-SY5Y cells when compared with non-treated cells. No modifications were seen in U87-MG cells. The increased A(2A)R mRNA and protein levels were accompanied by a loss of DNA methylation pattern in HeLa and SH-SY5Y cells, as measured with the SEQUENOM MassArray platform. The Aza treatment also reduced the affinity of a methyl-CpG-binding protein for ADORA2A by quantitative chromatin immunoprecipitation in HeLa cells. Interestingly, A(2A)R levels were reduced by S-adenosyl-l-methionine treatment in U87-MG and methyl-CpG-binding protein affinity was increased for ADORA2A by quantitative chromatin immunoprecipitation. Therefore, these results show for the first time that DNA methylation plays a role in ADORA2A transcription and, subsequently, in constitutive A(2A)R cell surface levels.
Buira
Sandra P
SP
Institut de Neuropatologia, Servei d'Anatomia Patològica, IDIBELL-Hospital Universitari de Bellvitge, L'Hospitalet de Llobregat, Spain.
Albasanz
José Luis
JL
Dentesano
Guido
G
Moreno
Jesús
J
Martín
Mairena
M
Ferrer
Isidre
I
Barrachina
Marta
M
D001374:Azacitidine / Q000494:pharmacology; D002460:Cell Line; D047369:Chromatin Immunoprecipitation / Q000379:methods; D019175:DNA Methylation / Q000187:drug effects / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D051783:Methyl-CpG-Binding Protein 2 / Q000378:metabolism; D009447:Neuroblastoma; D011485:Protein Binding / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D011869:Radioligand Assay / Q000379:methods; D043705:Receptor, Adenosine A2A / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D012436:S-Adenosylmethionine / Q000494:pharmacology; D013997:Time Factors; D014227:Triazines / Q000493:pharmacokinetics; D014230:Triazoles / Q000493:pharmacokinetics; D014316:Tritium / Q000493:pharmacokinetics
D004791:Enzyme Inhibitors; D051783:Methyl-CpG-Binding Protein 2; D012333:RNA, Messenger; D043705:Receptor, Adenosine A2A; D011956:Receptors, Cell Surface; D014227:Triazines; D014230:Triazoles; C097270:ZM 241385; D014316:Tritium; D012436:S-Adenosylmethionine; D001374:Azacitidine