Redundant regulation of urokinase plasminogen activator transcription by the two major isozymes of cAMP-dependent protein kinase.
The regulation of urokinase plasminogen activator (uPA) gene expression by the two major cAMP-dependent protein kinase isozymes was studied in SC115 mouse mammary carcinoma cells using the site-selective cAMP analog approach. SC115 cells expressed both type I and type II cAMP-dependent protein kinase holoenzyme (at a ratio of 2:3), and selective, partial activation of each holoenzyme could be demonstrated in vitro using appropriate combinations of cAMP analogs. When cells were exposed to the same analog combinations, uPA expression was upregulated 2- to 4-fold when either holoenzyme I or holoenzyme II was targeted. For comparison, a high concentration (1 mM) of 8-bromo-cAMP, an analog that does not discriminate between kinase isoforms, up-regulated uPA 10-fold. These findings suggest that there are two pathways of cAMP-dependent regulation of uPA, one mediated by holoenzyme I, the other by holoenzyme II, and that the end result of activation of each pathway is the same. Differences in the mechanism whereby each pathway regulates uPA were searched for but not found. Both pathways were shown to be dependent on catalytically active enzyme, to be potentiated by retinoic acid treatment, and to regulate uPA transcriptionally. The most likely interpretation of these findings is that uPA transcription is mediated solely by the action of the common catalytic subunit, regardless of whether it originated from holoenzyme I or holoenzyme II.
Mira-y-Lopez
R
R
Department of Medicine, Mount Sinai Medical School, City University of New York, New York 10029.
Jaramillo
S
S
Waxman
S
S
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D002999:Clone Cells; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D007700:Kinetics; D008325:Mammary Neoplasms, Experimental; D051379:Mice; D011494:Protein Kinases / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
D007527:Isoenzymes; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000242:Cyclic AMP; D011494:Protein Kinases; D014568:Urokinase-Type Plasminogen Activator