Differential regulation of hCG alpha and beta subunit mRNAs in JEG-3 choriocarcinoma cells by 8-bromo-cAMP.
The coordinate regulation of human chorionic gonadotropin (hCG) subunit synthesis by JEG-3 choriocarcinoma cells was studied at the pretranslational level. The responses of the hCG alpha and beta mRNAs were measured during stimulation with the potent cAMP analog 8-bromo-cAMP (8-Br-cAMP) using 32P-labeled hCG alpha and beta cDNA probes. The hCG alpha mRNA (850 bases) and beta mRNA (1050 bases) from JEG-3 cells were identical in size to that of their respective mRNAs from placenta, by Northern blot analysis. After 48 h of stimulation with 2 mM 8-Br-cAMP, production of immunoreactive alpha and beta subunits increased 25- and 52-fold, respectively; corresponding levels of the alpha and beta mRNAs increased 36- and 43-fold, respectively, in a dot blot hybridization assay. Total cellular protein, DNA content, and messenger RNA pools were not altered by treatment with 8-Br-cAMP. The temporal coordination of the expression of the hCG alpha- and beta-subunit genes was examined by comparing the time course of stimulation of the respective mRNAs and the production of immunoreactive subunits. The kinetic responses of the alpha and beta mRNAs differed: the increase in hCG alpha mRNA preceded the increase in hCG beta mRNA, while levels of free alpha subunit and intact hCG increased in parallel with the increase in beta mRNA. hCG alpha mRNA levels increased rapidly between 8 and 24 h after the addition of 8-Br-cAMP, and approached a plateau by 48 h. The levels of hCG beta mRNA increased steadily throughout the 8-48 h period. These results demonstrate that the cAMP analog 8-Br-cAMP differentially regulates hCG subunit biosynthesis in JEG-3 cells at a pretranslational level, and that the stimulation by 8-Br-cAMP in this system appears to be relatively selective for hCG subunits.
J
J
Burnside
S B
SB
Nagelberg
S S
SS
Lippman
B D
BD
Weintraub
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D002460:Cell Line; D002822:Choriocarcinoma / Q000378:metabolism*; D006063:Chorionic Gonadotropin / Q000235:genetics*; D018997:Chorionic Gonadotropin, beta Subunit, Human; D003545:Cysteine / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005260:Female; D015292:Glycoprotein Hormones, alpha Subunit; D006801:Humans; D007163:Immunosorbent Techniques; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D010446:Peptide Fragments / Q000235:genetics*; D010920:Placenta / Q000032:analysis; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis*
D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D015292:Glycoprotein Hormones, alpha Subunit; D010446:Peptide Fragments; D012333:RNA, Messenger; D015124:8-Bromo Cyclic Adenosine Monophosphate; D004247:DNA; D003545:Cysteine