Prolactin-mediated inhibition of 20alpha-hydroxysteroid dehydrogenase gene expression and the tyrosine kinase system.
The rat luteal 20alpha-hydroxysteroid dehydrogenase plays a key role at catabolizing progesterone and at decreasing the level of this steroid secreted by the ovaries. Throughout pregnancy and before parturition neither the mRNA nor the protein for this enzyme could be detected. In this investigation we set to examine whether PRL and PRL-like hormone from placental origin silence the expression of this gene and whether PRL action involves tyrosine kinase activity and/or de novo protein synthesis. The results revealed that PRL and PRL-like hormone from rat placental origin (rPL-1 and rPL-2), but not rat growth hormone, caused a rapid and profound inhibition of 20alpha-HSD mRNA expression in highly luteinized granulosa cells. Immunoprecipition and western blot analysis indicate that PRL-R associates with JAK2 and Stat5, and this association is increased within 30 seconds with PRL treatment. Although both JAK2 and Stat5 were phosphorylated on tyrosine upon PRL treatment, the PRL mediated inhibition of 20alpha-HSD was not reversed by either tyrosine kinase inhibitors, AG18 and genistein, but was largely reversed by the protein synthesis inhibitor cycloheximide. In summary, results of this investigation indicate that although PRL can activate the JAK2/Stat5 system in the corpus luteum, the down regulation of 20alpha-HSD mRNA by PRL does not appear to involve tyrosine kinase activity but depends on de novo synthesis of protein(s).
Zhong
L
L
Department of Physiology and Biophysics, University of Illinois at Chicago, 60612, USA.
Parmer
T G
TG
Robertson
M C
MC
Gibori
G
G
D015089:20-Hydroxysteroid Dehydrogenases / Q000096:biosynthesis*; D043607:20-alpha-Hydroxysteroid Dehydrogenase; D000818:Animals; D002478:Cells, Cultured; D003338:Corpus Luteum / Q000201:enzymology; D003513:Cycloheximide / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology*; D019833:Genistein; D006107:Granulosa Cells / Q000166:cytology / Q000201:enzymology*; D007529:Isoflavones / Q000494:pharmacology; D053614:Janus Kinase 2; D007700:Kinetics; D007743:Labor, Obstetric; D010920:Placenta / Q000502:physiology; D011247:Pregnancy; D011388:Prolactin / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D050797:STAT4 Transcription Factor; D015534:Trans-Activators / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D007529:Isoflavones; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D050797:STAT4 Transcription Factor; D015534:Trans-Activators; D011388:Prolactin; D003513:Cycloheximide; D019833:Genistein; D015089:20-Hydroxysteroid Dehydrogenases; D043607:20-alpha-Hydroxysteroid Dehydrogenase; D011505:Protein-Tyrosine Kinases; C507927:Jak2 protein, rat; D053614:Janus Kinase 2