Hilgers K F, Nagaraj S K, Karginova E A, Kazakova I G, Chevalier R L, Carey R M, Pentz E S, Gomez R A
Departments of Pediatrics and Internal Medicine, University of Virginia School of Medicine, Charlottesville, Virginia, USA.
Kidney Int. 1998 Nov;54(5):1444-54. doi: 10.1046/j.1523-1755.1998.00143.x.
We aimed to identify genes with kidney specific, developmentally regulated expression. Here we report the cDNA sequence and expression pattern of KS, a novel kidney-specific rat gene.
A partial cDNA was identified by differential display polymerase chain reaction (PCR) of a renal cell fraction enriched for proximal tubular and renin-expressing cells. Using the partial cDNA as a probe, a rat kidney cDNA library was screened. The full-length KS sequence was obtained by PCR amplification of cDNA ends. The expression pattern of KS was investigated by Northern blot. RNA was extracted from several organs of newborn and adult rats, as well as from the kidneys of rats with altered tubular function, that is, rats that had undergone unilateral nephrectomy, unilateral ureteral obstruction, neonatal losartan treatment, and the appropriate control animals. The expression of KS was also investigated in the kidneys of rats with spontaneous or renovascular hypertension.
The KS cDNA (2426 bp) contained one open reading frame encoding a predicted 572 amino acid protein. The derived peptide sequence displayed approximately 70% similarity to the hypertension-related SA gene product and approximately 50% similarity to prokaryotic and eukaryotic acetyl-CoA synthases (EC 6. 2.1.1). KS was expressed in the kidney and not in any other organ assayed. KS RNA was not detected in fetal and newborn rat kidney but became apparent after one week of postnatal life. Gene expression was downregulated in rat models of altered tubular function. KS expression was decreased in spontaneously hypertensive rats but not in renovascular hypertension.
KS, a novel rat gene, exhibits a unique tissue-specific expression exclusively in mature kidneys. The data suggest KS may encode an adenosine monophosphate binding enzyme.
我们旨在鉴定具有肾脏特异性、发育调控表达的基因。在此，我们报告一种新型肾脏特异性大鼠基因KS的cDNA序列及表达模式。
通过对富含近端小管和表达肾素细胞的肾细胞组分进行差异显示聚合酶链反应（PCR），鉴定出一个部分cDNA。以该部分cDNA为探针，筛选大鼠肾脏cDNA文库。通过cDNA末端的PCR扩增获得KS的全长序列。采用Northern印迹法研究KS的表达模式。从新生和成年大鼠的多个器官以及肾小管功能改变的大鼠（即接受单侧肾切除术、单侧输尿管梗阻、新生儿氯沙坦治疗的大鼠）及其相应对照动物的肾脏中提取RNA。还研究了自发性或肾血管性高血压大鼠肾脏中KS的表达。
KS cDNA（2426 bp）包含一个开放阅读框，编码一个预测的572个氨基酸的蛋白质。推导的肽序列与高血压相关的SA基因产物显示出约70%的相似性，与原核和真核乙酰辅酶A合成酶（EC 6.2.1.1）显示出约50%的相似性。KS在肾脏中表达，在其他检测的器官中不表达。在胎鼠和新生大鼠肾脏中未检测到KS RNA，但在出生后一周后变得明显。在肾小管功能改变的大鼠模型中基因表达下调。在自发性高血压大鼠中KS表达降低，但在肾血管性高血压中未降低。
KS是一种新型大鼠基因，仅在成熟肾脏中表现出独特的组织特异性表达。数据表明KS可能编码一种单磷酸腺苷结合酶。
