Tsai-Morris C H, Buczko E, Geng Y, Gamboa-Pinto A, Dufau M L
Section on Molecular Endocrinology, Endocrinology and Reproduction Research Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892-4510, USA.
J Biol Chem. 1996 Jun 14;271(24):14519-25. doi: 10.1074/jbc.271.24.14519.
Isolation and structural characterization of the rat corticotropin releasing factor receptor (CRFR) gene was performed to determine the exon/intron organization of the coding region and the potential for splice variants. The CRFR gene contains 13 exons and 12 introns, and the positions of the exon/intron junctions are similar to those of other Class II G protein-coupled receptor genes including the parathyroid hormone and glucagon receptors. The promoter resides within 593 base pairs of the initiation codon and the major transcriptional start site at nucleotide -238. This domain does not possess a TATA box but contains multiple Sp1 and AP-2 sites upstream and downstream of the major transcriptional start site. Intron junctions were identified in the extracellular, transmembrane (TM), and cytoplasmic (C) domains of the CRFR, giving the potential for differential signal transduction by splice variants. CRFR cDNAs derived from rat Leydig cell mRNA included the pituitary Form A, which spans exons 1-13, and two splice variants with deletion of exon 3 or exons 7, 11, and 12. An evolutionary link between the intronless TM/C module of the glycoprotein hormone receptors and the intron-containing TM/C module of the CRFR is suggested by the common position of the luteinizing hormone receptor Form D alternate acceptor splice site and the CRFR intron 12.
为了确定编码区的外显子/内含子组织以及剪接变体的可能性,对大鼠促肾上腺皮质激素释放因子受体(CRFR)基因进行了分离和结构表征。CRFR基因包含13个外显子和12个内含子,外显子/内含子连接位点的位置与其他II类G蛋白偶联受体基因(包括甲状旁腺激素和胰高血糖素受体)相似。启动子位于起始密码子的593个碱基对范围内,主要转录起始位点位于核苷酸-238处。该区域不具有TATA盒,但在主要转录起始位点的上游和下游包含多个Sp1和AP-2位点。在CRFR的细胞外、跨膜(TM)和细胞质(C)结构域中鉴定出内含子连接,这使得剪接变体有可能进行差异信号转导。源自大鼠睾丸间质细胞mRNA的CRFR cDNA包括跨越外显子1-13的垂体A型,以及缺失外显子3或外显子7、11和12的两种剪接变体。促黄体生成素受体D型交替受体剪接位点和CRFR内含子12的共同位置表明了糖蛋白激素受体的无内含子TM/C模块与CRFR的含内含子TM/C模块之间的进化联系。