Positive regulation of retinoic acid receptor alpha by protein kinase C and mitogen-activated protein kinase in sertoli cells.
Retinoic acid receptor alpha (RARalpha) is required for normal testis function. Similar to other steroid hormone receptors, RARalpha appears to undergo an activation process by which it translocates from the cytoplasm to the nucleus where it acts as a transcription factor. In this report, we demonstrate that RARalpha nuclear trafficking in Sertoli cells is positively regulated by phorbol-12-myristate-13-acetate-activated protein kinase C without the requirement of ligand, retinoic acid. Protein kinase C then stimulates the downstream mitogen-activated protein kinase, and the nuclear localization of RARalpha is dependent on activation of both kinases. The increase in RARalpha nuclear translocation is also coupled with enhanced transcriptional activity of RARalpha. This mechanism of RARalpha positive regulation is unique, different from that of its negative regulation, that has previously been shown to be dependent on cAMP-dependent protein kinase A and more importantly, dependent on its ligand. However, the mechanism by which retinoic acid positively influences the nuclear localization of RARalpha is not due to retinoic acid directly increasing protein kinase C or mitogen-activated protein kinase activities. Nonetheless, the positive influence of retinoic acid is also dependent on these two kinases as determined by inhibitor studies. These results suggest two mechanisms for RARalpha activation in Sertoli cells: one involving only the two kinases, the other involving both the ligand and the two kinases. These regulatory mechanisms for RARalpha activation, both positive and negative, may be critical for the proper function of RARalpha in the testis.
Braun
Kirt W
KW
School of Molecular Biosciences, Center for Reproductive Biology, Washington State University, Pullman, WA 99164, USA.
Vo
My-Nuong
MN
Kim
Kwan Hee
KH
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D008024:Ligands; D008156:Luciferases / Q000378:metabolism; D008297:Male; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D009687:Nuclear Proteins / Q000378:metabolism; D011493:Protein Kinase C / Q000502:physiology*; D051381:Rats; D018168:Receptors, Retinoic Acid / Q000096:biosynthesis*; D000072482:Retinoic Acid Receptor alpha; D012176:Retinoids / Q000494:pharmacology; D012708:Sertoli Cells / Q000201:enzymology / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000235:genetics
D008024:Ligands; D009687:Nuclear Proteins; C000607072:Rara protein, mouse; C000607073:Rara protein, rat; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha; D012176:Retinoids; D008156:Luciferases; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases; D013755:Tetradecanoylphorbol Acetate