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DNA methylation regulates adenosine A 2A receptor cell surface expression levels
Author(s) -
Buira Sandra P.,
Albasanz José Luis,
Dentesano Guido,
Moreno Jesús,
Martín Mairena,
Ferrer Isidre,
Barrachina Marta
Publication year - 2010
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2009.06538.x
Subject(s) - chromatin immunoprecipitation , hela , dna methylation , microbiology and biotechnology , immunoprecipitation , biology , methylation , receptor , cpg site , chromatin , gene expression , cell , promoter , dna , gene , biochemistry
J. Neurochem. (2010) 112 , 1273–1285. Abstract 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 μM 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.