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调控元件的特征在于嘌呤介导的和细胞类型特异性的基因转录。

A regulatory element is characterized by purine-mediated and cell-type-specific gene transcription.

作者信息

Walsh M J, Sanchez-Pozo A, Leleiko N S

机构信息

Department of Pediatrics, Mount Sinai School of Medicine, New York, New York 10029.

出版信息

Mol Cell Biol. 1990 Aug;10(8):4356-64. doi: 10.1128/mcb.10.8.4356-4364.1990.

Abstract

Purines and purine nucleotides were found to affect transcription of the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene in whole nuclei isolated from intestinal mucosa of adult rats fed a purine- and purine nucleotide-free diet. Nuclear run-on transcription assays, performed on whole nuclei from different tissues and cell types, identified an intestine-specific decrease in the overall incorporation of [alpha-32P]UTP in HPRT transcripts from intestinal epithelial cell nuclei when exogenous purines or purine nucleotides were omitted from either the diet or culture medium. Using a 990-base-pair genomic fragment that contains the 5'-flanking region from the HPRT gene, we generated plasmid constructs with deletions, transfected the DNA into various cell types, and assayed for chloramphenicol acetyltransferase (CAT) reporter activity in vitro. We determined that an element upstream from the putative transcriptional start site is necessary to maintain the regulatory response to purine and nucleotide levels in cultured intestinal epithelial cells. These results were tissue and cell type specific and suggest that in the absence of exogenous purines, the presence of specific factors influences transcriptional initiation of HPRT. This information provides evidence for a mechanism by which the intestinal epithelium, which has been reported to lack constitutive levels of de novo purine nucleotide biosynthetic activity, could maintain and regulate the salvage of purines and nucleotides necessary for its high rate of cell and protein turnover during fluctuating nutritional and physiological conditions. Furthermore, this information may provide more insight into regulation of the broad class of genes recognized by their lack of TATA and CCAAT box consensus sequences within the region proximal to the promoter.

摘要

在给成年大鼠喂食不含嘌呤和嘌呤核苷酸的饮食后,从其肠黏膜分离出的完整细胞核中发现,嘌呤和嘌呤核苷酸会影响次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(HPRT)基因的转录。对来自不同组织和细胞类型的完整细胞核进行的核转录延伸分析表明,当从饮食或培养基中去除外源性嘌呤或嘌呤核苷酸时,肠上皮细胞核中HPRT转录本中[α - 32P]UTP的总体掺入量出现肠道特异性下降。利用一个包含HPRT基因5'侧翼区域的990个碱基对的基因组片段,我们构建了缺失突变的质粒构建体,将DNA转染到各种细胞类型中,并在体外检测氯霉素乙酰转移酶(CAT)报告基因的活性。我们确定,假定转录起始位点上游的一个元件对于维持培养的肠上皮细胞中对嘌呤和核苷酸水平的调节反应是必需的。这些结果具有组织和细胞类型特异性,表明在没有外源性嘌呤的情况下,特定因子的存在会影响HPRT的转录起始。这一信息为一种机制提供了证据,据报道,肠上皮细胞缺乏从头合成嘌呤核苷酸的组成性活性,在营养和生理条件波动时,该机制可维持和调节其高细胞和蛋白质周转率所需的嘌呤和核苷酸的补救合成。此外,这一信息可能会为对一类因其启动子近端区域缺乏TATA和CCAAT框共有序列而被识别的基因的调控提供更多见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d21e/360986/7342b3fbb0a8/molcellb00044-0512-a.jpg

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