Histone deacetylase inhibitors decrease reelin promoter methylation in vitro.
We investigated the effects of agents that induce reelin mRNA expression in vitro on the methylation status of the human reelin promoter in neural progenitor cells (NT2). NT2 cells were treated with the histone deacetylase inhibitors, trichostatin A (TSA) and valproic acid (VPA), and the methylation inhibitor aza-2'-deoxycytidine (AZA) for various times. All three drugs reduced the methylation profile of the reelin promoter relative to untreated cells. The acetylation status of histones H3 and H4 increased following treatment with VPA and TSA at times as short as 15 min following treatment; a result consistent with the reported mode of action of these drugs. Chromatin immunoprecipitation experiments showed that these changes were accompanied by changes occurring at the level of the reelin promoter as well. Interestingly, AZA decreased reelin promoter methylation without concomittantly increasing histone acetylation. In fact, after prolonged treatments with AZA, the acetylation status of histones H3 and H4 decreased relative to untreated cells. We also observed a trend towards reduced methylated H3 after 18 h treatment with TSA and VPA. Our data indicate that while TSA and VPA act to increase histone acetylation and reduce promoter methylation, AZA acts only to decrease the amount of reelin promoter methylation.
Mitchell
Colin P
CP
The Psychiatric Institute, Department of Psychiatry, University of Illinois at Chicago, Illinois 60612, USA.
Chen
Ying
Y
Kundakovic
Marija
M
Costa
Erminio
E
Grayson
Dennis R
DR
D015816:Cell Adhesion Molecules, Neuronal / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D019175:DNA Methylation / Q000187:drug effects*; D016326:Extracellular Matrix Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic / Q000187:drug effects* / Q000235:genetics*; D011480:Protease Inhibitors / Q000494:pharmacology*; D000091042:Reelin Protein; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism
D015816:Cell Adhesion Molecules, Neuronal; D016326:Extracellular Matrix Proteins; D056572:Histone Deacetylase Inhibitors; D009419:Nerve Tissue Proteins; D011480:Protease Inhibitors; D000091042:Reelin Protein; C000717127:RELN protein, human; D012697:Serine Endopeptidases; D006655:Histone Deacetylases