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

被引:27
作者
Seki, Y
Ikeda, S
Kiyohara, H
Ayabe, H
Seki, T
Matsui, H
机构
[1] Okayama Univ Sci, Dept Biochem, Okayama 7000005, Japan
[2] Okayama Univ, Grad Sch Med & Dent, Dept Physiol, Okayama 7008558, Japan
[3] Univ Florida, Coll Med, Dept Physiol, Gainesville, FL 32610 USA
关键词
Pasteurella haemolytica A1 glycoprotease; APEX nuclease; luciferase assay; CCAAT box; cap site hunting;
D O I
10.1016/S0378-1119(02)00429-8
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
We performed cDNA and genomic cloning, sequencing and promoter analysis of the putative human 0-sialoglycoprotein endopeptidase gene OSGEP (a homologue of gep, a Pasteurella haemolytica A 1 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. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:101 / 108
页数:8
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