Sequencing analysis of a putative human O-sialoglycoprotein endopeptidase gene (OSGEP) and analysis of a bidirectional promoter between the OSGEP and APEX genes.
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.
Seki
Yuichi
Y
Department of Physiology, Okayama University Graduate School of Medicine and Dentistry, 700-8558, Okayama, Japan.
Ikeda
Shogo
S
Kiyohara
Hiroyuki
H
Ayabe
Hiroaki
H
Seki
Tsugio
T
Matsui
Hideki
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D019757:Carbon-Oxygen Lyases / Q000235:genetics*; D004247:DNA / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D005091:Exons; D005260:Female; D015870:Gene Expression; D005796:Genes / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D018076:DNA, Complementary; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA; D008156:Luciferases; D008666:Metalloendopeptidases; C070738:O-sialoglycoprotein endopeptidase; D019757:Carbon-Oxygen Lyases; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase