THE CLONING OF THE HUMAN FOLLICLE-STIMULATING-HORMONE RECEPTOR AND ITS EXPRESSION IN COS-7, CHO, AND Y-1 CELLS
THE CLONING OF THE HUMAN FOLLICLE-STIMULATING-HORMONE RECEPTOR AND ITS EXPRESSION IN COS-7, CHO, AND Y-1 CELLS
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DOI：
10.1016/0303-7207(92)90220-z
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发表时间：
1992-11-01
影响因子：
4.1
通讯作者：
CHAPPEL, SC
中科院分区：
文献类型：
--
作者：
KELTON, CA;CHENG, SVY;CHAPPEL, SC
从人睾丸 cDNA 文库中分离出卵泡刺激素 (FSH) 受体克隆。 cDNA克隆的表征表明，长开放阅读框的DNA和预测氨基酸序列与先前发表的人卵巢FSH受体序列（Minegish等人（1991）Biochem.Biophys.Res.Commun.175, 1125-1130）有七个核苷酸和五个氨基酸不同。还鉴定并表征了人类 FSH 受体剪接变体。全长人类 FSH 受体 cDNA 被设计用于在 COS-7、CHO 和 Y-1 细胞中表达。在 COS-7 细胞的瞬时转染和 Y-1 细胞的稳定转染中，只有当 FSH 受体转录单位的 5' 非翻译区包含内含子时，才会发生有效的 FSH 受体 mRNA 积累和 FSH 响应细胞系的分离。稳定转染 FSH 受体的 Y-1 细胞通过四舍五入和合成更多量的黄体酮来响应 FSH 处理。无论是否存在异源内含子，均分离出稳定转染的 CHO 细胞系，该细胞系通过合成增加量的 cAMP 来响应 FSH。 FSH 反应性 CHO 和 Y-1 细胞系可能适合开发更好的体外 FSH 生物测定法。这些细胞还构成了人类 FSH 受体蛋白的便利来源，用于放射受体测定和受体-配体相互作用的研究。
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.