Jameson L, Chin W W, Hollenberg A N, Chang A S, Habener J F
J Biol Chem. 1984 Dec 25;259(24):15474-80.
The nucleotide sequence of the gene encoding the beta-subunit of rat luteinizing hormone (LH beta) has been determined from a genomic DNA fragment cloned in lambda phage Charon 4A. Blot hybridization of restriction enzyme digests of rat genomic DNA indicates that the gene is present in a single copy. The transcriptional unit is 0.98 kilobase in size and contains three exons interrupted by two introns of 245 and 225 base pairs (bp). The locations of the exon/intron junctions at amino acid codons -16/-15 and +41/+42 have been conserved between the rat LH beta gene and the related genes, human LH beta and human chorionic gonadotropin beta. Using S1 nuclease mapping and oligonucleotide-primed reverse transcription of ovariectomized rat pituitary mRNA, the start of transcription was determined to be 7 bp upstream from the start of translation. Characteristic promoter elements are present in the 5'-flanking region of the gene, including the Goldberg-Hogness sequence, TATAAA, 31 bp, and the consensus CAAT box sequence, 167 bp upstream from the start of transcription, respectively. Within the proximal 200 bp flanking the 5'-region of the transcriptional unit, there is strong homology between the rat and human LH beta genes, suggesting that these regions include sequences which may be important for regulation of gene expression. Isolation and characterization of the rat LH beta gene further defines the evolution of glycoprotein hormone genes and will facilitate the study of cellular and molecular mechanisms which regulate LH beta gene expression.
已从克隆于λ噬菌体Charon 4A的基因组DNA片段中确定了编码大鼠促黄体激素(LHβ)β亚基的基因的核苷酸序列。对大鼠基因组DNA进行限制性酶切消化后的印迹杂交表明该基因以单拷贝形式存在。转录单位大小为0.98千碱基,包含三个外显子,被两个分别为245和225个碱基对(bp)的内含子打断。大鼠LHβ基因与相关基因人LHβ和人绒毛膜促性腺激素β之间,外显子/内含子连接在氨基酸密码子-16/-15和+41/+42处的位置是保守的。通过S1核酸酶图谱分析以及对去卵巢大鼠垂体mRNA进行寡核苷酸引发的逆转录,确定转录起始位点在翻译起始位点上游7 bp处。在该基因的5'侧翼区域存在特征性的启动子元件,分别包括戈德堡-霍格尼斯序列TATAAA(位于转录起始位点上游31 bp处)和共有CAAT框序列(位于转录起始位点上游167 bp处)。在转录单位5'区域侧翼的近端200 bp范围内,大鼠和人LHβ基因之间存在高度同源性,这表明这些区域包含可能对基因表达调控很重要的序列。大鼠LHβ基因的分离和特性鉴定进一步明确了糖蛋白激素基因的进化,并将有助于研究调节LHβ基因表达的细胞和分子机制。