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首页> 外文期刊>Zoological Science >Expression and genomic organization of a medaka fish novel membrane form of guanylyl cyclase/orphan receptor
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Expression and genomic organization of a medaka fish novel membrane form of guanylyl cyclase/orphan receptor

机译：aka鸟新型膜形式鸟苷酸环化酶/孤儿受体的表达和基因组组织

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摘要

A novel membrane guanylyl cyclase (membrane GC), OIGC8, was identified in the medaka fish Oryzias latipes by the isolation of full-length cDNA (4958 bp) and genomic DNA (14.3 kbp) clones. Phylogenetic analysis indicated that OIGC8 does not belong in any known vertebrate membrane GC subfamily OIGC8 consists of an extracellular domain (214 residues), a transmembrane segment (19 residues), and an intracellular protein kinase-like domain (284 residues) and a cyclase catalytic domain (228 residues), although the extracellular domain is about half the length (around 450 residues) of other known vertebrate membrane GCs. OIGC8 transiently expressed in COS-7 cells exhibited only basal guanylyl cyclase activity. None of the known ligands (rat ANP, BNP, CNP, and C-ANF) and various medaka fish tissue extracts, which activated OIGC1, OIGC2, and OIGC7 differentially, stimulated basal activity, suggesting that OIGC8 is an orphan receptor. The OIGC8 gene consists of 24 exons and exists as a single copy on the medaka fish genome. Northern blot hybridization showed that a 5 kb-OIGC8 mRNA was expressed in the kidney and the testis at a high level and a 3.3 kb-OIGC8 mRNA was expressed only in the brain. The RNase protection, RNA Ligase-Mediated Rapid Amplification of cDNA Ends (RLM-RACE), and reverse transcription-polymerase chain reaction (RT-PCR) analyses demonstrated that the 3.3 kb-OIGC8 mRNA detected in the brain is transcribed from the second transcription initiation site, and contains an intron at the position prior to the catalytic domain, the translation product of which appears to be a protein lacking the cyclase catalytic domain.
机译：通过分离全长cDNA（4958 bp）和基因组DNA（14.3 kbp）克隆，在in鱼Oryzias latipes中鉴定了一种新型膜鸟苷酰环化酶（膜GC）OIGC8。系统发育分析表明，OIGC8不属于任何已知的脊椎动物膜GC亚家族，OIGC8由细胞外域（214个残基），跨膜片段（19个残基）和细胞内蛋白激酶样结构域（284个残基）和环化酶催化组成（228个残基），尽管胞外域的长度约为其他已知脊椎动物膜GC的一半（约450个残基）。在COS-7细胞中瞬时表达的OIGC8仅表现出基础鸟苷基环化酶活性。已知的配体（大鼠ANP，BNP，CNP和C-ANF）和各种medaka鱼组织提取物均不能差异地激活OIGC1，OIGC2和OIGC7，不会刺激其基础活性，这表明OIGC8是一个孤儿受体。 OIGC8基因由24个外显子组成，并在medaka鱼基因组中单拷贝存在。 Northern印迹杂交显示，在肾脏和睾丸中高水平表达了5 kb-OIGC8 mRNA，仅在大脑中表达了3.3 kb-OIGC8 mRNA。 RNase保护，RNA连接酶介导的cDNA末端快速扩增（RLM-RACE）和逆转录聚合酶链反应（RT-PCR）分析表明，从第二次转录中转录出在大脑中检测到的3.3 kb-OIGC8 mRNA。起始位点，并在催化结构域之前的位置含有一个内含子，其翻译产物似乎是缺少环化酶催化结构域的蛋白质。

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