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首页> 外文期刊>The biochemical journal >Murine cystathionine γ-lyase: complete cDNA and genomic sequences, promoter activity, tissue distribution and developmental expression
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Murine cystathionine γ-lyase: complete cDNA and genomic sequences, promoter activity, tissue distribution and developmental expression

机译:鼠胱硫醚γ-裂合酶:完整的cDNA和基因组序列,启动子活性,组织分布和发育表达

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pCystathionine γ-lyase (CSE) is the last key enzyme in the trans-sulphuration pathway for biosynthesis of cysteine from methionine. Cysteine could be provided through diet; however, CSE has been shown to be important for the adequate supply of cysteine to synthesize glutathione, a major intracellular antioxidant. With a view to determining physiological roles of CSE in mice, we report the sequence of a complete mouse CSE cDNA along with its associated genomic structure, generation of specific polyclonal antibodies, and the tissue distribution and developmental expression patterns of CSE in mice. A 1.8 kb full-length cDNA containing an open reading frame of 1197 bp, which encodes a 43.6 kDa protein, was isolated from adult mouse kidney. A 35 kb mouse genomic fragment was obtained by λ genomic library screening. It contained promoter regions, 12 exons, ranging in size from 53 to 579 bp, spanning over 30 kb, and exon/intron boundaries that were conserved with rat and human iCSE/i. The GC-rich core promoter contained canonical TATA and CAAT motifs, and several transcription factor-binding consensus sequences. The iCSE/i transcript, protein and enzymic activity were detected in liver, kidney, and, at much lower levels, in small intestine and stomach of both rats and mice. In developing mouse liver and kidney, the expression levels of CSE protein and activity gradually increased with age until reaching their peak value at 3 weeks of age, following which the expression levels in liver remained constant, whereas those in kidney decreased significantly. Immunohistochemical analyses revealed predominant CSE expression in hepatocytes and kidney cortical tubuli. These results suggest important physiological roles for CSE in mice./p
机译:半胱氨酸硫氨酸γ-裂解酶(CSE)是反硫途径中从蛋氨酸生物合成半胱氨酸中的最后一个关键酶。半胱氨酸可以通过饮食来提供;然而,CSE已被证明对于充足供应半胱氨酸以合成谷胱甘肽(一种主要的细胞内抗氧化剂)很重要。为了确定CSE在小鼠中的生理作用,我们报告了完整的小鼠CSE cDNA序列及其相关的基因组结构,特异性多克隆抗体的产生以及小鼠中CSE的组织分布和发育表达模式。从成年小鼠肾脏中分离出一个1.8kb的全长cDNA,该cDNA含有一个1197bp的开放阅读框,该编码框编码43.6kDa的蛋白质。通过λ基因组文库筛选获得了35kb的小鼠基因组片段。它包含启动子区域,12个外显子,大小从53到579bp,跨度超过30kb,以及外显子/内含子边界,与大鼠和人CSE保守。富含GC的核心启动子包含规范的TATA和CAAT图案,以及几个转录因子结合共有序列。在大鼠和小鼠的肝脏,肾脏以及小肠和胃中均检测到 CSE 转录本,蛋白质和酶活性。在发育中的小鼠肝脏和肾脏中,CSE蛋白和活性的表达水平随着年龄的增长而逐渐增加,直到在3周龄时达到峰值,此后肝脏中的表达水平保持恒定,而肾脏中的表达水平则明显下降。免疫组织化学分析显示,CSE在肝细胞和肾皮质小管中占主导。这些结果表明CSE在小鼠中具有重要的生理作用。

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