Seimiya M, Kusakabe T, Suzuki N
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060, Japan.
J Biol Chem. 1997 Sep 12;272(37):23407-17. doi: 10.1074/jbc.272.37.23407.
Three cDNAs (OlGC3, OlGC4, and OlGC5) encoding membrane guanylyl cyclases were isolated from a medaka (Oryzias latipes) eye cDNA library. An open reading frame for OlGC3 predicted a protein of 1057 amino acids, and those for OlGC4 and OlGC5, 1134 and 1151, respectively. These proteins consist of an apparent signal peptide (21 residues for OlGC3, 50 residues for OlGC4, and 48 residues for OlGC5) and a single transmembrane domain that divides the protein into an amino-terminal extracellular domain and a carboxyl-terminal intracellular domain that further divides into a kinase-like domain and a cyclase catalytic domain. Phylogenetic analysis with amino acid sequences of OlGC3, OlGC4, and OlGC5, as well as those of other membrane guanylyl cyclases, indicated that OlGC3, OlGC4, and OlGC5 are members of the sensory organ-specific guanylyl cyclase family. Reverse transcription-polymerase chain reaction and Northern blot analyses demonstrated that OlGC3, OlGC4, and OlGC5 transcripts are present in the eye, which contains more cGMP than the other organs. In addition to being expressed in the eye, OlGC3 transcripts are also present in the brain, heart, liver, pancreas, and ovary, while OlGC4 is present in the liver and OlGC5 in the heart. Reverse transcription-polymerase chain reaction analysis with RNA from unfertilized eggs and embryos showed that OlGC3 and OlGC5 are expressed both maternally and zygotically, while OlGC4 is expressed only zygotically, and that the zygotic expression of these three genes is differentially activated. These results suggest a structural and functional diversity of sensory organ-specific guanylyl cyclases in vertebrates.
从青鳉(Oryzias latipes)眼cDNA文库中分离出三个编码膜鸟苷酸环化酶的cDNA(OlGC3、OlGC4和OlGC5)。OlGC3的开放阅读框预测编码一个1057个氨基酸的蛋白质,OlGC4和OlGC5的开放阅读框分别预测编码1134和1151个氨基酸的蛋白质。这些蛋白质均包含一个明显的信号肽(OlGC3为21个残基,OlGC4为50个残基,OlGC5为48个残基)和一个单一跨膜结构域,该跨膜结构域将蛋白质分为氨基末端胞外结构域和羧基末端胞内结构域,后者又进一步分为激酶样结构域和环化酶催化结构域。对OlGC3、OlGC4和OlGC5以及其他膜鸟苷酸环化酶的氨基酸序列进行系统发育分析表明,OlGC3、OlGC4和OlGC5是感觉器官特异性鸟苷酸环化酶家族的成员。逆转录-聚合酶链反应和Northern印迹分析表明,OlGC3、OlGC4和OlGC5转录本存在于眼中,眼内的环磷酸鸟苷(cGMP)含量高于其他器官。除了在眼中表达外,OlGC3转录本也存在于脑、心脏、肝脏、胰腺和卵巢中,而OlGC4存在于肝脏中,OlGC5存在于心脏中。对未受精卵和胚胎的RNA进行逆转录-聚合酶链反应分析表明,OlGC3和OlGC5在母源和合子阶段均有表达,而OlGC4仅在合子阶段表达,且这三个基因的合子表达被差异激活。这些结果表明脊椎动物中感觉器官特异性鸟苷酸环化酶存在结构和功能多样性。