Molecular cloning, genomic organization and developmental regulation of a novel receptor from Drosophila melanogaster structurally related to gonadotropin-releasing hormone receptors from vertebrates
Molecular cloning, genomic organization and developmental regulation of a novel receptor from Drosophila melanogaster structurally related to gonadotropin-releasing hormone receptors from vertebrates
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DOI：
10.1006/bbrc.1998.9230
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发表时间：
1998-08-28
影响因子：
3.1
通讯作者：
Grimmelikhuijzen, CJP
中科院分区：
文献类型：
--
作者：
Hauser, F;Sondergaard, L;Grimmelikhuijzen, CJP
在用编码G蛋白偶联受体跨膜区的序列对伯克利果蝇基因组计划的数据库进行筛选后，我们发现果蝇可能含有编码一种受体的基因，该受体在结构上与脊椎动物的促性腺激素释放激素（GnRH）受体相关。使用聚合酶链反应，果蝇cDNA作为模板，和寡核苷酸探针编码该基因的假定外显子，我们能够克隆该受体的cDNA编码。该受体的跨膜区与鲶鱼的跨膜区有36%的氨基酸同源性，与大鼠GnRH受体的跨膜区有31%的氨基酸同源性。果蝇受体基因含有6个内含子，而大鼠基因含有2个;果蝇基因中的一个内含子与大鼠基因中的一个内含子位于完全相同的位置，并具有相同的内含子相位，表明果蝇和哺乳动物GnRH受体基因在进化上相关。北方印迹分析表明，果蝇受体基因在幼虫发育过程中进行性表达，在3龄幼虫阶段显著最大。蛹含有少量的受体mRNA，而成年果蝇含有较高的水平，雄性果蝇的受体mRNA比雌性果蝇多约5倍。Southern印迹分析表明，果蝇只含有一个拷贝的受体基因，位于2号染色体的27 A2-B1位置。本文首次报道了无脊椎动物GnRH受体家族成员的分子克隆。(C)北京：科学出版社.
After screening the data base of the Berkeley Drosophila Genome Project with a sequence coding for the transmembrane region of a G protein-coupled receptor, we found that Drosophila might contain a gene coding for a receptor that is structurally related to the Gonadotropin-Releasing Hormone (GnRH) receptors from vertebrates. Using the polymerase chain reaction, with Drosophila cDNA as a template, and oligonucleotide probes coding for the presumed exons of this gene, we were able to clone the cDNA coding for this receptor. The transmembrane region of the receptor shows 36% amino acid residue identity with the transmembrane region of the catfish and 31% amino acid residue identity with that of the rat GnRH receptor. The Drosophila receptor gene contains six introns, whereas the rat gene contains two; one intron in the Drosophila gene occurs at exactly the same position and has the same intron phasing as one intron in the rat gene, suggesting that the Drosophila and mammalian GnRH receptor genes are evolutionarily related. Northern blot analyses show that the Drosophila receptor gene is progressively expressed during larval development with a prominent maximum at the 3rd instar larval stage. Pupae contain low amounts of receptor mRNA, while adult flies contain higher levels, with males having about five times more receptor mRNA than female files. Southern blot analyses show that Drosophila contains only one copy of the receptor gene, which is located at position 27A2-B1 of chromosome 2. This paper is the first report on the molecular cloning of a member of the GnRH receptor family from invertebrates. (C) 1998 Academic Press.