The ratPDE3/IVd phosphodiesterase gene codes for multiple proteins differentially activated by cAMP-dependent protein kinase.
Several mRNAs coding for a cAMP-specific phosphodiesterase (ratPDE3/IVd) with divergent 5' regions have been detected in mammalian cells. To determine the physiological significance of these differences, the expression of these mRNAs and the properties of the corresponding proteins were investigated. At least three mRNA species derived from the ratPDE3/IVd gene (ratPDE3.1, ratPDE3.2, and ratPDE3.3 mRNAs) are present in Sertoli and thyroid cells and in brain. Expression of ratPDE3.1 and ratPDE3.2 but not ratPDE3.3 mRNAs was dependent on hormone stimulation. The ratPDE3.2 and ratPDE3.3 mRNA variants were translated into polypeptides with immunochemical and biochemical properties identical to the native cAMP phosphodiesterases (PDEs) found in the Sertoli cell and thyroid FRTL-5 cells. Incubation of the recombinant PDEs with the catalytic subunit of the cAMP-dependent protein kinase in a cell-free system caused the phosphorylation and activation of the ratPDE3.3 protein variant. Under the same experimental conditions, ratPDE3.1 and ratPDE3.2 protein variants were neither phosphorylated nor activated by the cAMP-dependent protein kinase. Similar results were obtained by stimulating cells expressing the three recombinant PDE variants with dibutyryl cAMP. These findings demonstrate that the ratPDE3/IVd gene codes for PDE forms subject to different regulations.
Department of Gynecology and Obstetrics, Stanford University, Medical Center, California 94305-5317.
C
C
Sette
E
E
Vicini
M
M
Conti
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D003994:Bucladesine / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D017931:DNA Primers; D018076:DNA, Complementary / Q000302:isolation & purification; D015971:Gene Expression Regulation, Enzymologic; D014644:Genetic Variation; D007527:Isoenzymes / Q000096:biosynthesis; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D010727:Phosphoric Diester Hydrolases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D012708:Sertoli Cells / Q000201:enzymology*; D013961:Thyroid Gland / Q000201:enzymology; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003994:Bucladesine; D017868:Cyclic AMP-Dependent Protein Kinases; D010727:Phosphoric Diester Hydrolases