Production of three distinct mRNAs of 150 kDa oxygen-regulated protein (ORP150) by alternative promoters: preferential induction of one species under stress conditions.
150 kDa oxygen-regulated protein (ORP150) is one of the endoplasmic reticulum (ER)-resident stress proteins. We have cloned and sequenced the entire human ORP150 gene covering over 15-kb. Analyses of transcription initiation sites and transcriptional regulatory sequences revealed that at least three distinct mRNA species were produced by alternative promoters: two of them starting from alternative exon 1 (1A or 1B), and the third one starting from exon 2, six nucleotides upstream of the first AUG initiation codon. Among them, the transcript that begins with exon 1B was preferentially induced by hypoxia or tunicamycin treatment. A cis-acting segment involved in the stress-dependent induction was found at the 5'-end of exon 1A, which could account for the selective induction of the transcription from exon 1B. Furthermore, in vitro analyses of translation of the third mRNA suggested the constitutive expression of the cytosolic ORP150 due to the lack of the signal peptide resulting from differential translation initiation.
Kaneda
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S
HSP Research Institute, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan.
Yura
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T
Yanagi
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H
D001483:Base Sequence; D015152:Blotting, Northern; D015687:Cell Hypoxia; D004587:Electrophoresis, Agar Gel; D005809:Genes, Regulator / Q000235:genetics*; D015698:Genomic Library; D018840:HSP70 Heat-Shock Proteins; D006801:Humans; D015151:Immunoblotting; D008156:Luciferases / Q000378:metabolism; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism; D013312:Stress, Physiological / Q000235:genetics*; D014407:Tumor Cells, Cultured
D018840:HSP70 Heat-Shock Proteins; D011506:Proteins; D012333:RNA, Messenger; C050087:oxygen-regulated proteins; D008156:Luciferases; D015719:Single-Strand Specific DNA and RNA Endonucleases