The cloning of the human follicle stimulating hormone receptor and its expression in COS-7, CHO, and Y-1 cells.
Follicle stimulating hormone (FSH) receptor clones were isolated from a human testis cDNA library. Characterization of the cDNA clones showed that the DNA and predicted amino acid sequences of the long open reading frame differed from a previously published human ovarian FSH receptor sequence (Minegish et al. (1991) Biochem. Biophys. Res. Commun. 175, 1125-1130) by seven nucleotides and five amino acids. A human FSH receptor splice variant was also identified and characterized. A full-length human FSH receptor cDNA was engineered for expression in COS-7, CHO, and Y-1 cells. In transient transfections of COS-7 cells and stable transfections of Y-1 cells, efficient FSH receptor mRNA accumulation and isolation of FSH-responsive cell lines occurred only when an intron was included in the 5' untranslated region of the FSH receptor transcription unit. Y-1 cells stably transfected with the FSH receptor responded to FSH treatment by rounding up and by synthesizing increased amounts of progesterone. Stably transfected CHO cell lines, which responded to FSH by synthesizing increased amounts of cAMP, were isolated irrespective of the presence of the heterologous intron. The FSH-responsive CHO and Y-1 cell lines may be suitable for the development of better in vitro FSH bioassays. These cells also constitute a convenient source of human FSH receptor protein for use in radioreceptor assays and in studies of receptor-ligand interactions.
Kelton
C A
CA
Ares Advanced Technology, Inc., Randolph, MA 02368.
Cheng
S V
SV
Nugent
N P
NP
Schweickhardt
R L
RL
Rosenthal
J L
JL
Overton
S A
SA
Wands
G D
GD
Kuzeja
J B
JB
Luchette
C A
CA
Chappel
S C
SC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D015723:Gene Library; D014644:Genetic Variation; D006801:Humans; D007438:Introns; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010053:Ovary / Q000737:chemistry; D011374:Progesterone / Q000096:biosynthesis; D051381:Rats; D011962:Receptors, FSH / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D012708:Sertoli Cells / Q000737:chemistry; D013737:Testis / Q000737:chemistry*; D014162:Transfection; D014407:Tumor Cells, Cultured
D011962:Receptors, FSH; D011993:Recombinant Fusion Proteins; D011374:Progesterone; D004247:DNA