Phosphodiesterase 2 negatively regulates adenosine-induced transcription of the tyrosine hydroxylase gene in PC12 rat pheochromocytoma cells.
Adenosine induces expression of the tyrosine hydroxylase (TH) gene in PC12 cells. However, it is suggested that atrial natriuretic peptide (ANP) inhibits expression of this gene. Using real-time PCR and luciferase reporter assays we found that ANP significantly decreases the adenosine-induced transcription of the TH gene. Results of measurements of cyclic nucleotide concentrations indicated that ANP-induced accumulation of cGMP inhibits the adenosine-induced increase in cAMP level. Using selective phosphodiesterase 2 (PDE2) inhibitors and a synthetic cGMP analog activating PDE2, we found that PDE2 is involved in coupling the ANP-triggered signal to the cAMP metabolism. We have established that ANP-induced elevated levels of cGMP as well as cGMP analog stimulate hydrolytic activity of PDE2, leading to inhibition of adenosine-induced transcription of the TH gene. We conclude that ANP mediates negative regulation of TH gene expression via stimulation of PDE2-dependent cAMP breakdown in PC12 cells.
Laboratory of Signaling Proteins, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland. Electronic address: edyta.makuch@iitd.pan.wroc.pl.
Edyta
E
Makuch
Laboratory of Signaling Proteins, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland.
Marianna
M
Kuropatwa
Laboratory of Signaling Proteins, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland.
Ewa
E
Kurowska
Laboratory of Biomedical Chemistry "Neolek", Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland.
Jaroslaw
J
Ciekot
Laboratory of Experimental Anticancer Therapy "Neolek", Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland.
Dagmara
D
Klopotowska
Laboratory of Signaling Proteins, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland.
Janusz
J
Matuszyk
D000241:Adenosine / Q000494:pharmacology*; D000310:Adrenal Gland Neoplasms / Q000201:enzymology* / Q000235:genetics*; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D054678:Cyclic Nucleotide Phosphodiesterases, Type 2 / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D008954:Models, Biological; D016716:PC12 Cells; D010673:Pheochromocytoma / Q000201:enzymology / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects*; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics* / Q000378:metabolism
D017362:Cyclic AMP Response Element-Binding Protein; D012333:RNA, Messenger; D009320:Atrial Natriuretic Factor; D000242:Cyclic AMP; D014446:Tyrosine 3-Monooxygenase; D054678:Cyclic Nucleotide Phosphodiesterases, Type 2; C517278:Pde2a protein, rat; D006152:Cyclic GMP; D000241:Adenosine
Adenosine; Atrial natriuretic peptide (ANP); Phosphodiesterase 2 (PDE2); Rat pheochromocytoma PC12 cells; Tyrosine hydroxylase