Atger M, Misrahi M, Sar S, Le Flem L, Dessen P, Milgrom E
Unité de Recherches Hormones et Reproduction, INSERM U.135, Le Kremlin Bicêtre, France.
Mol Cell Endocrinol. 1995 Jun;111(2):113-23. doi: 10.1016/0303-7207(95)03557-n.
The complete organization of the human luteinizing hormone-choriogonadotropin (LH/CG) receptor (LH/CGR) gene and the structure of 1591 bp of its 5' flanking region have been determined. This gene spans over 70 kbp and contains 11 exons. The first ten exons and part of the last exon encode the extracellular domain of the receptor while the transmembrane and intracellular domains are encoded by the remaining part of the last exon. The gene encodes a 701 amino acids long preprotein, contrary to a previous report of 699 amino acids. Primer extension experiments and polymerase chain reaction (PCR) mapping allowed definition of the transcription initiation site, which is located 1085 bp upstream from the initiation codon. The 5' non-coding region is thus unusually long. The promoter region which is different from the murine LH/CG receptor promoter, contains two putative TATA boxes at positions -34 and -47 and a CAAT box consensus sequence at position -89. A consensus sequence corresponding to a cAMP responsive element is found at position -697. Seven API consensus sequences are also found in the 5' flanking region of the gene. Southern blot experiments demonstrated an informative biallelic polymorphism within the human LH/CG receptor gene locus using BglII endonuclease. The cloning of the human LH/CGR gene and the determination of the organization and structure of its 5' flanking region allow the study of its hormonal, developmental and tissue-specific regulation. Primers and PCR conditions are described for the direct genomic sequencing of all the exons of the gene. This information should facilitate the study of pathological mutations of the receptor.
已确定人促黄体生成素 - 绒毛膜促性腺激素（LH/CG）受体（LH/CGR）基因的完整结构及其5'侧翼区1591 bp的结构。该基因跨度超过70 kbp，包含11个外显子。前十个外显子和最后一个外显子的一部分编码受体的细胞外结构域，而跨膜和细胞内结构域由最后一个外显子的其余部分编码。该基因编码一个701个氨基酸长的前体蛋白，与之前报道的699个氨基酸不同。引物延伸实验和聚合酶链反应（PCR）定位确定了转录起始位点，该位点位于起始密码子上游1085 bp处。因此，5'非编码区异常长。与小鼠LH/CG受体启动子不同的启动子区域，在 -34和 -47位置含有两个假定的TATA盒，在 -89位置含有一个CAAT盒共有序列。在 -697位置发现了一个与cAMP反应元件相对应的共有序列。在该基因的5'侧翼区还发现了七个AP1共有序列。Southern印迹实验表明，使用BglII核酸内切酶在人LH/CG受体基因位点内存在一个信息丰富的双等位基因多态性。人LH/CGR基因的克隆及其5'侧翼区的组织和结构的确定有助于研究其激素、发育和组织特异性调节。描述了用于该基因所有外显子直接基因组测序的引物和PCR条件。这些信息应有助于研究该受体的病理突变。
