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一个假定的人类O-唾液酸糖蛋白内肽酶基因(OSGEP)的测序分析以及OSGEP与APEX基因之间双向启动子的分析。

Sequencing analysis of a putative human O-sialoglycoprotein endopeptidase gene (OSGEP) and analysis of a bidirectional promoter between the OSGEP and APEX genes.

作者信息

Seki Yuichi, Ikeda Shogo, Kiyohara Hiroyuki, Ayabe Hiroaki, Seki Tsugio, Matsui Hideki

机构信息

Department of Physiology, Okayama University Graduate School of Medicine and Dentistry, 700-8558, Okayama, Japan.

出版信息

Gene. 2002 Feb 20;285(1-2):101-8. doi: 10.1016/s0378-1119(02)00429-8.

Abstract

We performed cDNA and genomic cloning, sequencing and promoter analysis of the putative human O-sialoglycoprotein endopeptidase gene OSGEP (a homologue of gcp, a Pasteurella haemolytica A1 glycoprotease). The cloned OSGEP cDNA is 1311 nucleotides long, and encodes a protein consisting of 335 amino acids with predicted molecular mass of 36.4 kDa. The amino acid sequence of OSGEP showed 29.7% identity with that of P. haemolytica glycoprotease. The OSGEP gene is 7.75 kb long, consists of 11 exons and 10 introns, and lies immediately adjacent to the APEX gene (which encodes APEX nuclease, a multifunctional DNA repair enzyme) in 5'-to-5' orientation. The promoter region of the OSGEP gene lacks the typical TATA box, but has putative regulatory elements in the CpG island. Northern blot analysis showed ubiquitous expression of the OSGEP gene in several tissues, and we observed similarities in expression patterns between OSGEP and APEX. In order to study the regulation of OSGEP gene expression, we analyzed the OSGEP promoter region by luciferase assay using HeLa cells. A functional region required for full transcription activity was narrowed down to a 23 bp region containing a CCAAT box. It has been reported that this CCAAT box promotes basal transcription in the APEX direction. We thus conclude that a bidirectional promoter containing a CCAAT box regulates transcription of both the OSGEP and APEX genes.

摘要

我们对假定的人类O-唾液酸糖蛋白内肽酶基因OSGEP(溶血巴斯德菌A1糖蛋白酶gcp的同源物)进行了cDNA和基因组克隆、测序及启动子分析。克隆的OSGEP cDNA长1311个核苷酸,编码一个由335个氨基酸组成的蛋白质,预测分子量为36.4 kDa。OSGEP的氨基酸序列与溶血巴斯德菌糖蛋白酶的氨基酸序列有29.7%的同一性。OSGEP基因长7.75 kb,由11个外显子和10个内含子组成,以5'至5'方向紧邻APEX基因(该基因编码APEX核酸酶,一种多功能DNA修复酶)。OSGEP基因的启动子区域缺乏典型的TATA盒,但在CpG岛中有假定的调控元件。Northern印迹分析显示OSGEP基因在多个组织中普遍表达,并且我们观察到OSGEP和APEX之间的表达模式相似。为了研究OSGEP基因表达的调控,我们使用HeLa细胞通过荧光素酶测定法分析了OSGEP启动子区域。完全转录活性所需的功能区域被缩小到一个包含CCAAT盒的23 bp区域。据报道,这个CCAAT盒在APEX方向促进基础转录。因此我们得出结论,一个包含CCAAT盒的双向启动子调节OSGEP和APEX基因的转录。

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