ISOLATION AND CHARACTERIZATION OF THE GENE ENCODING THE BETA-SUBUNIT OF RAT FOLLICLE-STIMULATING-HORMONE
ISOLATION AND CHARACTERIZATION OF THE GENE ENCODING THE BETA-SUBUNIT OF RAT FOLLICLE-STIMULATING-HORMONE
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DOI：
10.1089/dna.1.1989.8.339
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发表时间：
1989-06-01
期刊：
影响因子：
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通讯作者：
CHIN, WW
中科院分区：
文献类型：
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作者：
GHARIB, SD;ROY, A;CHIN, WW
促卵泡激素（FSH）是一种垂体糖蛋白激素，由两个不同的亚基α和β组成。和β，由不同的基因编码我们已经分离了编码β-通过与牛FSH-β过滤杂交筛选大鼠基因组DNA文库，cDNA。大鼠基因组DNA的Southern印迹分析表明，存在FSH-β的单拷贝。每个单倍体基因组的基因数。大鼠FSH-β的核苷酸序列基因的克隆，并推导了该亚基的氨基酸序列。该基因由三个外显子和两个内含子组成。预测的氨基酸序列表明，有一个20个氨基酸的信号肽，其次是一个成熟的蛋白质的110个氨基酸残基。外显子I、II和III分别为36、187和1221 bp。内含子1（640 bp）中断5“-非翻译（UT）区（61 bp），内含子2（约1 kb）中断氨基酸残基+34和+35之间的编码区。氨基酸序列与人和牛FSH-β的氨基酸序列的比较亚基揭示了80%的相似性，这两个物种。由外显子II编码的氨基酸的“CAGY”序列或Cys-Ala-Gly-Tyr四联体存在于每一个其它糖蛋白β中。到目前为止测序的亚基在大鼠FSH-β中改变，其中Ala残基被Glu取代。引物延伸分析表明，有一个单一的转录起始位点。测定5“-侧翼区（1 kb）的核苷酸序列，并与牛和人基因中相应区域的核苷酸序列进行比较。该分析揭示了在大鼠FSH-β的5 ″-侧翼区中有三个区域。与牛和人基因中的区域显示大于80%序列相似性的基因。这些区域中的第二个也与大鼠α-SMA的5“-侧翼区域内的区段共享相似性。和LH β-亚基基因大鼠FSH-β的表征基因的表达将使进一步研究大鼠FSH的调控使用体外系统。
Follicle-stimulating hormone (FSH) is a pituitary glycoprotein hormone that is comprised of two dissimilar subunits, .alpha. and .beta., encoded by separate genes. We have isolated the gene encoding the .beta.-subunit of rat FSH by screening a rat genomic DNA library by filter hybridization with bovine FSH-.beta. cDNA. Southern blot analysis of rat genomic DNA suggests that there is a single copy of the FSH-.beta. gene per haploid genome in the rat. The nucleotide sequence of the rat FSH-.beta. gene was determined and the amino acid sequence of the subunit was deduced. The gene is composed of three exons and two introns. The predicted amino acid sequence reveals that there is a 20-amino-acid signal peptide followed by a mature protein of 110 amino acid residues. Exons, I, II, and III are 36, 187, and 1221 bp, respectively. Intron 1 (640 bp) interrupts the 5''-untranslated (UT) region (61 pb) and intron 2 (approximately 1 kb) interrupts the coding region between amino acid residues +34 and +35. Comparison of the amino acid sequence to those of the human and bovine FSH-.beta. subunits reveals 80% similarity to both species. The "CAGY" sequence, or the Cys-Ala-Gly-Tyr quartet of amino acids encoded by exon II is present in every other glycoprotein .beta.-subunit sequenced thus far, is altered in rat FSH-.beta., with the Ala residue replaced by Glu. Primer extension analysis demonstrated that there is a single transcriptional start site. The nucleotide sequence of the 5''-flanking region (1 kb) was determined and compared to the nucleotide sequences of corresponding regions in the bovine and human genes. This analysis revealed that there are three regions in the 5''-flanking region of the rat FSH-.beta. gene that display greater than 80% sequence similarity to regions in the bovine and human genes. The second of these regions also shares similarity to segments within the 5''-flanking regions of the rat .alpha.-and LH.beta.-subunit genes. The characterization of the rat FSH-.beta. gene will enable further study of the regulation of rat FSH using in vitro systems.