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首页> 外文期刊>American Journal of Physiology >Characterization of the 5'-flanking region of OK cell type II Na-Pi cotransporter gene.
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Characterization of the 5'-flanking region of OK cell type II Na-Pi cotransporter gene.

机译：OK细胞II型Na-Pi共转运蛋白基因的5'侧翼区的表征。

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摘要

The renal type II Na-Pi cotransport is the rate-limiting step in proximal tubular phosphate (Pi) reabsorption. Among the different "proximal tubular" cell lines, this transporter seem only to be expressed in opossum kidney cells (OK cells). We have isolated the 5'-flanking region of the ok-Npt2 gene (OK cell type II Na-Pi cotransporter) including exons 1-3 and containing a TFIID site (TATA box), a GCCAAT site, an AP1 site, and two microsatellite GGAA repeats. Major transcription initiation sites were determined by primer extension and rapid amplification of 5' cDNA ends (5'-RACE). A 327-bp fragment containing the TFIID and GCAAT element was driving the downstream luciferase reporter gene in homologous transfection assays. Slightly reduced promoter activity was observed with a 198-bp fragment containing the GCAAT element; shorter fragments were without activity. Promoter activity (327-bp fragment) could also be observed in transfections into HeLa cells but not in U937 human macrophage cells, MCT mouse kidney cortex cells, and MDCK cells. Different "physiological" stimuli known to be associated with altered proximal tubular Na-Pi cotransport activity are without effect on transcriptional activity in above homologous transfection experiments.
机译：肾脏II型Na-Pi共转运是近端管状磷酸盐（Pi）重吸收的限速步骤。在不同的“近端管状”细胞系中，这种转运蛋白似乎仅在负鼠肾细胞（OK细胞）中表达。我们已经分离出ok-Npt2基因的5'侧翼区域（OK细胞II型Na-Pi共转运蛋白），包括外显子1-3，并包含一个TFIID位点（TATA框），一个GCCAAT位点，一个AP1位点和两个微卫星GGAA重复。通过引物延伸和5'cDNA末端（5'-RACE）的快速扩增来确定主要的转录起始位点。在同源转染试验中，含有TFIID和GCAAT元件的327 bp片段正在驱动下游的荧光素酶报道基因。用含有GCAAT元件的198bp片段观察到启动子活性略有降低。较短的片段没有活性。在转染到HeLa细胞中也观察到了启动子活性（327bp片段），但在U937人巨噬细胞，MCT小鼠肾皮质细胞和MDCK细胞中没有观察到。在上述同源转染实验中，已知与近端肾小管Na-Pi共转运活性改变相关的不同“生理”刺激对转录活性没有影响。

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