The effects of sodium butyrate on transcription are mediated through activation of a protein phosphatase.
In this study we have investigated the molecular mechanism by which sodium butyrate modulates gene expression when added to cultured cells. As a model system we used hepatoma tissue culture cells in which sodium butyrate treatment increases histone H1(0) mRNA level and decreases c-myc mRNA level. Because we observed that stimulation of histone H1(0) gene expression could take place in the absence of protein neosynthesis, we hypothesized that sodium butyrate induced a post-translational modification of a factor involved in the transcription process. Using different types of well known kinase and phosphatase inhibitors, we studied the implication of kinase or phosphatase activity in this pathway. Interestingly, cell treatment with potent serine-threonine-phosphatase inhibitors, calyculin A or okadaic acid, prevented the regulation of both histone H1(0) and c-myc gene expressions by sodium butyrate. On the other hand, the tyrosine phosphatase inhibitor, vanadate, or the protein kinase C inhibitor, staurosporine, did not significantly modify sodium butyrate effects. Using protein phosphatase 1 and 2A for in vitro assays, we found a 45% increase of phosphatase activity after cell treatment by sodium butyrate, possibly due to a protein phosphatase 1-type protein phosphatase. These data strongly suggest that signaling pathway(s) triggered by sodium butyrate to modulate gene expression involve(s) a serine-threonine-phosphatase activity.
Cuisset
L
L
Laboratoire de Biologie Moléculaire des Cellules Eucaryotes, I.C.G. M-EA 1501, Université Paris V, René Descartes, Faculté de Médecine Cochin Port-Royal, 24 rue du Faubourg Saint-Jacques, 75014 Paris, France.
Tichonicky
L
L
Jaffray
P
P
Delpech
M
M
D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016259:Genes, myc; D006657:Histones / Q000235:genetics; D008387:Marine Toxins; D010080:Oxazoles / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054645:Protein Phosphatase 1; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014407:Tumor Cells, Cultured
D002087:Butyrates; D004791:Enzyme Inhibitors; D006657:Histones; D008387:Marine Toxins; D010080:Oxazoles; D012333:RNA, Messenger; D020148:Butyric Acid; C059041:calyculin A; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1