Involvement of an inhibitory G-protein in the signal transduction pathway of maturation-inducing hormone (17 alpha,20 beta-dihydroxy-4-pregnen-3-one) action in rainbow trout (Oncorhynchus mykiss) oocytes.
To investigate the mechanism by which the hormone 17 alpha,20 beta-dihydroxy-4-pregnen-3-one(17 alpha,20 beta-DP) acts on a receptor on the external surface of rainbow trout oocytes to induce maturation, the interaction between 17 alpha,20 beta-DP receptors and G-proteins was examined. Pertussis toxin (PT) catalyzed the ADP ribosylation of a 40-kDa protein in crude membranes from rainbow trout postvitellogenic oocytes, and cholera toxin (CT) labeled several proteins, including a major protein with an apparent molecular weight of 43 kDa. The 40-kDa protein was recognized by an antibody against the alpha subunit of inhibitory G-proteins (Gi), whereas the 43-kDa protein was recognized by an antibody against the alpha subunit of stimulatory G-proteins (Gs). Treating the membrane fraction with 17 alpha,20 beta-DP decreased the PT-catalyzed ADP ribosylation of the 40-kDa protein. In contrast, there was no significant change in the CT-catalyzed ribosylation of the 43-kDa protein after exposure to 17 alpha,20 beta-DP. The specific binding of 17 alpha,20 beta-DP to membrane fractions was decreased by PT. 17 alpha,20 beta-DP binding was also inhibited by nonhydrolyzable GTP analogs such as guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and guanylylimidodiphosphate (GppNHp), but not by either ATP or guanosine 5'-O-(2-thiodiphosphate) (GDP beta S). Scatchard analysis revealed that GppNHp induced a 3.8-fold increase in the dissociation constant without a significant change in the number of binding sites, suggesting that the GppNHp-induced decrease in 17 alpha,20 beta-DP binding is due to the decrease in binding affinity between 17 alpha,20 beta-DP and its receptors. We conclude that the PT-sensitive Gi is involved in the signal transduction pathway of 17 alpha,20 beta-DP in rainbow trout oocytes.
Yoshikuni
M
M
Department of Developmental Biology, National Institute for Basic Biology, Okazaki, Japan.
Nagahama
Y
Y
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002772:Cholera Toxin / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000502:physiology*; D006160:Guanosine Triphosphate / Q000502:physiology; D006908:Hydroxyprogesterones / Q000378:metabolism / Q000494:pharmacology*; D008540:Meiosis / Q000187:drug effects*; D017686:Oncorhynchus mykiss / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*; D037342:Pertussis Toxin; D011956:Receptors, Cell Surface / Q000378:metabolism; D015398:Signal Transduction; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D006908:Hydroxyprogesterones; D011956:Receptors, Cell Surface; D010566:Virulence Factors, Bordetella; C007433:17,20-dihydroxy-4-pregnen-3-one; D000246:Adenosine Diphosphate Ribose; D006160:Guanosine Triphosphate; D002772:Cholera Toxin; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases