Iio Kaori, Nakauchi Mina, Yamagami Sayaka, Tsutsumi Makiko, Hori Hiroshi, Naruse Kiyoshi, Mitani Hiroshi, Shima Akihiro, Suzuki Norio
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
Comp Biochem Physiol B Biochem Mol Biol. 2005 Apr;140(4):569-78. doi: 10.1016/j.cbpc.2004.12.006. Epub 2005 Jan 13.
A novel membrane guanylyl cyclase (GC), OlGC9, was identified in the intestine of the medaka fish Oryzias latipes by the isolation of a full-length cDNA clone (3783 bp). Phylogenetic analysis indicated that OlGC9 belongs in the enterotoxin/guanylin receptor membrane GC subfamily. The nucleotide and deduced amino acid sequences of OlGC9 were highly homologous to those of OlGC6, another enterotoxin/guanylin receptor membrane GC in medaka fish. Linkage analysis of the medaka fish chromosome demonstrated that the OlGC9 gene was mapped to LG8, which distinguishes it from the OlGC6 gene. Determination of the cGMP concentrations in COS-7 cells expressed with OlGC9 indicated that Escherichia coli heat-stable enterotoxin (STa) stimulated the activity of OlGC9 in a concentration-dependent manner, although it did not activate the OlGC6 expressed in the COS-7 cells. The 5'-flanking region of the OlGC9 gene important for its transcription was partially determined using both CACO-2 cells and COS-1 cells, and was not found to be conserved with respect to either the mammalian GC-C gene or the OlGC6 gene.
通过分离一个全长cDNA克隆(3783 bp),在青鳉鱼(Oryzias latipes)的肠道中鉴定出一种新型膜鸟苷酸环化酶(GC),即OlGC9。系统发育分析表明,OlGC9属于肠毒素/鸟苷蛋白受体膜GC亚家族。OlGC9的核苷酸序列和推导的氨基酸序列与青鳉鱼中另一种肠毒素/鸟苷蛋白受体膜GC即OlGC6的高度同源。青鳉鱼染色体的连锁分析表明,OlGC9基因定位于LG8,这使其与OlGC6基因区分开来。对用OlGC9表达的COS-7细胞中cGMP浓度的测定表明,大肠杆菌热稳定肠毒素(STa)以浓度依赖的方式刺激OlGC9的活性,尽管它不激活在COS-7细胞中表达的OlGC。使用CACO-2细胞和COS-1细胞部分确定了对OlGC9基因转录重要的5'侧翼区域,发现其在哺乳动物GC-C基因或OlGC6基因方面均不保守。