Protein tyrosine phosphatases are involved in LH/chorionic gonadotropin and 8Br-cAMP regulation of steroidogenesis and StAR protein levels in MA-10 Leydig cells.
The LH signal transduction pathway features the activation of protein tyrosine phosphatases (PTPs) as one of the components of a cascade that includes other well characterized events such as cAMP-dependent protein kinase A (PKA) activation. Moreover, the action of PTPs is required to increase the rate-limiting step in steroid biosynthesis, namely the cAMP-regulated transfer of cholesterol to the inner mitochondrial membrane. Since both PKA activity and steroidogenic acute regulatory (StAR) protein induction are obligatory steps in this transfer of cholesterol, the present study was performed to investigate the role of PTPs in the regulation of PKA activity and StAR expression in response to LH/chorionic gonadotropin (CG) and 8Br-cAMP in MA-10 cells. While the exposure of MA-10 cells to the PTP inhibitor, phenylarsine oxide (PAO), did not modify PKA activity, it partially inhibited the effect of human CG and cAMP analog on StAR protein levels. Time-course studies demonstrated that PAO inhibited cAMP induction of StAR protein and mRNA. At 30 min, the effect on cAMP-stimulated StAR protein levels was a 35% inhibition, progressing to up to 90% inhibition at 120 min of stimulation. The maximal inhibitory effect on cAMP-induced StAR mRNA level was obtained at 60 min (85%). In summary, these results demonstrate that inhibition of PTP activity affected both StAR protein and mRNA synthesis and suggest that the activity of hormone-regulated PTPs is a requirement in the LH signaling cascade that results in the up-regulation of StAR protein and, subsequently, increased steroid synthesis.
Paz
C
C
Department of Biochemistry, School of Medicine, University of Buenos Aires, Paraguay 2155, 5th, (C1121ABG) Buenos Aires, Argentina.
Cornejo Maciel
F
F
Maloberti
P
P
Walsh
L P
LP
Stocco
D M
DM
Podestá
E J
EJ
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D000728:Androgens / Q000096:biosynthesis*; D000818:Animals; D001152:Arsenicals / Q000494:pharmacology; D002784:Cholesterol / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000235:genetics; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D015398:Signal Transduction; D013997:Time Factors; D014407:Tumor Cells, Cultured; D000098682:Steroidogenic Acute Regulatory Protein
D000728:Androgens; D001152:Arsenicals; D006063:Chorionic Gonadotropin; D004791:Enzyme Inhibitors; D010750:Phosphoproteins; D012333:RNA, Messenger; D000098682:Steroidogenic Acute Regulatory Protein; C029341:oxophenylarsine; D015124:8-Bromo Cyclic Adenosine Monophosphate; D002784:Cholesterol; D017868:Cyclic AMP-Dependent Protein Kinases; D017027:Protein Tyrosine Phosphatases