Functional properties of the rat phosphatase 1alpha promoter.
The aim of this study was to investigate the functional properties of the promoter of the protein phosphatase 1alpha catalytic subunit. Luciferase plasmids with different fragments of the rat catalytic subunit of the protein phosphatase 1alpha promoter ranging from -3.7 kbp to -59 bp were transiently transfected into cells by the calcium-phosphate precipitation method. The promoter activity was determined in the absence and presence of inotropic agents which influencing the cAMP-depending pathway. The basal transcriptional activity decreased at fragment -124 bp and shorter fragments. To identify regions of regulatory importance we investigated the cAMP-dependent influence on the transcriptional activity. Stimulation of the complete promoter region with forskolin (1-100 microM) for 6 h led to a concentration-dependent decrease of transcriptional activity. Moreover, regions shorter than 3.7 kbp were inhibited by forskolin (10 microM). Short time stimulation (10 min) with forskolin (10 microM) increased the transcriptional activity of only the 3.7 kbp fragment. The effects were antagonized by Rp-cAMPS, a specific antagonist of protein kinase A, indicating cAMP-dependent effects. The results provide evidence for cAMP-dependent regulation of the protein phosphatase 1alpha promotor.
Linck
B
B
Institut für Pharmakologie und Toxikologie, Universität Münster, Germany.
Boknik
P
P
Knapp
J
J
Kikuchi
K
K
Lüss
H
H
Müller
F U
FU
Nomoto
K
K
Neumann
J
J
Schmitz
W
W
D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D003994:Bucladesine / Q000494:pharmacology; D020134:Catalytic Domain; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017930:Genes, Reporter; D006801:Humans; D008969:Molecular Sequence Data; D009206:Myocardium / Q000166:cytology; D010749:Phosphoprotein Phosphatases / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013873:Thionucleotides / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D004791:Enzyme Inhibitors; D011993:Recombinant Fusion Proteins; D013873:Thionucleotides; D005576:Colforsin; C016957:adenosine-3',5'-cyclic phosphorothioate; D003994:Bucladesine; D000242:Cyclic AMP; D010749:Phosphoprotein Phosphatases