Cooperation between phosphorylation and acetylation processes in transcriptional control.
We previously reported that the activation of the M promoter of the human choline acetyltransferase (ChAT) gene by butyrate and trapoxin in transfected CHP126 cells is blocked by PD98059, a specific mitogen-activated protein kinase kinase (MEK) inhibitor (E. Espinos and M. J. Weber, Mol. Brain Res. 56:118-124, 1998). We now report that the transcriptional effects of histone deacetylase inhibitors are mediated by an H7-sensitive serine/threonine protein kinase. Activation of the ChAT promoter by butyrate and trapoxin was blocked by 50 microM H7 in both transient- and stable-transfection assays. Overexpression of p300, a coactivator protein endowed with histone acetyltransferase activity, stimulated the ChAT promoter and had a synergistic effect on butyrate treatment. These effects were blocked by H7 and by overexpressed adenovirus E1A 12S protein. Moreover, both H7 and PD98059 suppressed the activation of the Rous sarcoma virus (RSV) and simian virus 40 promoters by butyrate in transfection experiments. Similarly, the induction of the cellular histone H1(0) gene by butyrate in CHP126 cells was blocked by H7 and by PD98059. Previous data (L. Cuisset, L. Tichonicky, P. Jaffray, and M. Delpech, J. Biol. Chem. 272:24148-24153, 1997) showed that the induction of the H1(0) gene by butyrate is blocked by okadaic acid, an inhibitor of protein phosphatases. We now show that the activation of the ChAT and RSV promoters by butyrate in transfected CHP126 cells is also blocked by 200 nM okadaic acid. Western blotting and in vivo metabolic labeling experiments showed that butyrate has a biphasic effect on histone H3 phosphorylation, i.e., depression for up to 16 h followed by stimulation. The data thus strongly suggest that the transcriptional effects of histone deacetylase inhibitors are mediated through the activation of MEK1 and of an H7-sensitive protein kinase in addition to protein phosphatases.
Espinos
E
E
Laboratoire de Biologie Moléculaire Eucaryote, CNRS UPR 9006, 31062 Toulouse Cedex, France.
Le Van Thaï
A
A
Pomiès
C
C
Weber
M J
MJ
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D000107:Acetylation; D000123:Acetyltransferases / Q000235:genetics; D017500:Adenovirus E1A Proteins / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002087:Butyrates / Q000494:pharmacology; D002453:Cell Cycle / Q000235:genetics; D018797:Cell Cycle Proteins / Q000235:genetics; D002460:Cell Line; D002795:Choline O-Acetyltransferase / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D051548:Histone Acetyltransferases; D056572:Histone Deacetylase Inhibitors; D006657:Histones / Q000235:genetics; D006801:Humans; D019319:Okadaic Acid / Q000494:pharmacology; D010455:Peptides; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D014157:Transcription Factors; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000123:Acetyltransferases; D017500:Adenovirus E1A Proteins; D000900:Anti-Bacterial Agents; D002087:Butyrates; D018797:Cell Cycle Proteins; D002795:Choline O-Acetyltransferase; D004791:Enzyme Inhibitors; D005419:Flavonoids; D051548:Histone Acetyltransferases; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D019319:Okadaic Acid; D010455:Peptides; D017346:Protein Serine-Threonine Kinases; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C067070:trapoxin A; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one