Human N‐Acetylglucosaminyltransferase III Gene is Transcribed from Multiple Promoters

We have isolated cDNA clones for the human N‐acetylglucosaminyltransferase III (GlcNAc‐transferase III) gene. Two of them, H15 and H20, contain 5′ non‐coding regions that are totally different from each other except for 8 bp adjacent to the putative initiation codon. Analysis of one of the genomic c...

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Veröffentlicht in:European journal of biochemistry 1996-06, Vol.238 (3), p.853-861
Hauptverfasser: Koyama, Nobuto, Miyoshi, Eiji, Ihara, Yoshito, Kang, Rujun, Nishikawa, Atsushi, Taniguchi, Naoyuki
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container_issue 3
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container_title European journal of biochemistry
container_volume 238
creator Koyama, Nobuto
Miyoshi, Eiji
Ihara, Yoshito
Kang, Rujun
Nishikawa, Atsushi
Taniguchi, Naoyuki
description We have isolated cDNA clones for the human N‐acetylglucosaminyltransferase III (GlcNAc‐transferase III) gene. Two of them, H15 and H20, contain 5′ non‐coding regions that are totally different from each other except for 8 bp adjacent to the putative initiation codon. Analysis of one of the genomic cosmid clones containing the GlcNAc–transferase III coding region, Hug3, revealed that the 5′ non‐coding regions of H15 and H20 contain two and one exons, respectively, in addition to the exon containing the coding region (exon 1). These have arisen as the result of alternative splicing. The transcription‐initiation sites were determined by primer‐extension analysis and 5′‐rapid amplification of cDNA ends (RACE). Both H15‐specific and H20‐specific primers gave cDNAs longer than those expected from the lengths of H15 and H20, and a primer complementary to the region around the intron/exon junction near the putative initiation codon also gave distinct signals. Promoter activities of the 5′‐flanking regions of H15, H20 and exon 1 were measured in a human hepatoblastoma cell line, HuH‐6 cells by luciferase assays. The 5′‐flanking region of exon 1 was the most active, whilst that of H15 was several times less active, and that of H20 was inactive. Our study suggests that multiple promoters of the GlcNAc‐transferase III gene contribute to the complex regulation of this gene.
doi_str_mv 10.1111/j.1432-1033.1996.0853w.x
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Two of them, H15 and H20, contain 5′ non‐coding regions that are totally different from each other except for 8 bp adjacent to the putative initiation codon. Analysis of one of the genomic cosmid clones containing the GlcNAc–transferase III coding region, Hug3, revealed that the 5′ non‐coding regions of H15 and H20 contain two and one exons, respectively, in addition to the exon containing the coding region (exon 1). These have arisen as the result of alternative splicing. The transcription‐initiation sites were determined by primer‐extension analysis and 5′‐rapid amplification of cDNA ends (RACE). Both H15‐specific and H20‐specific primers gave cDNAs longer than those expected from the lengths of H15 and H20, and a primer complementary to the region around the intron/exon junction near the putative initiation codon also gave distinct signals. Promoter activities of the 5′‐flanking regions of H15, H20 and exon 1 were measured in a human hepatoblastoma cell line, HuH‐6 cells by luciferase assays. The 5′‐flanking region of exon 1 was the most active, whilst that of H15 was several times less active, and that of H20 was inactive. 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Two of them, H15 and H20, contain 5′ non‐coding regions that are totally different from each other except for 8 bp adjacent to the putative initiation codon. Analysis of one of the genomic cosmid clones containing the GlcNAc–transferase III coding region, Hug3, revealed that the 5′ non‐coding regions of H15 and H20 contain two and one exons, respectively, in addition to the exon containing the coding region (exon 1). These have arisen as the result of alternative splicing. The transcription‐initiation sites were determined by primer‐extension analysis and 5′‐rapid amplification of cDNA ends (RACE). Both H15‐specific and H20‐specific primers gave cDNAs longer than those expected from the lengths of H15 and H20, and a primer complementary to the region around the intron/exon junction near the putative initiation codon also gave distinct signals. Promoter activities of the 5′‐flanking regions of H15, H20 and exon 1 were measured in a human hepatoblastoma cell line, HuH‐6 cells by luciferase assays. The 5′‐flanking region of exon 1 was the most active, whilst that of H15 was several times less active, and that of H20 was inactive. Our study suggests that multiple promoters of the GlcNAc‐transferase III gene contribute to the complex regulation of this gene.</description><subject>Alternative Splicing</subject><subject>Base Sequence</subject><subject>Chromosome Mapping</subject><subject>DNA Primers</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>gene promoter system</subject><subject>genomic structure</subject><subject>Hepatoblastoma - genetics</subject><subject>Hepatoblastoma - pathology</subject><subject>Humans</subject><subject>luciferase assay</subject><subject>Molecular Sequence Data</subject><subject>N-Acetylglucosaminyltransferases - biosynthesis</subject><subject>N-Acetylglucosaminyltransferases - genetics</subject><subject>N‐acetylglucosaminyltransferase III</subject><subject>N‐linked oligosaccharide</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Promoter Regions, Genetic</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Sequence Analysis, DNA</subject><subject>Transcription, Genetic</subject><subject>Tumor Cells, Cultured</subject><issn>0014-2956</issn><issn>1432-1033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkM1q3DAQx0VpSLdpH6GgU292R5IlWZdCGvKxkCaBpuxRaOVx8SLbW8km2Vseoc_YJ4mdXXJt5zIf___MwI8QyiBnU3zZ5KwQPGMgRM6MUTmUUjzkj2_I4lV4SxYArMi4keodeZ_SBgCUUfqYHJcalDKwIKursXUdvfn79OfU47ALv8Lo--TaptuFIbou1RhdQrpcLukldkibRO_nuY_NGitax76l38cwNNuA9G7q-gFj-kCOahcSfjzkE_Lz4vz-7Cq7vr1cnp1eZ74oilXmvREGasmdMr4Qiq-hMqAqLZkGzx1qxrDmEqpCSODMMI_caMe51EIoJk7I5_3dbex_j5gG2zbJYwiuw35MVpdMcwbyn0YmS8250JOx3Bt97FOKWNttbFoXd5aBneHbjZ0Z25mxneHbF_j2cVr9dPgxrlusXhcPtCf9615_aALu_vuuvTj_9mMqV-IZEmuT7w</recordid><startdate>199606</startdate><enddate>199606</enddate><creator>Koyama, Nobuto</creator><creator>Miyoshi, Eiji</creator><creator>Ihara, Yoshito</creator><creator>Kang, Rujun</creator><creator>Nishikawa, Atsushi</creator><creator>Taniguchi, Naoyuki</creator><general>Blackwell Science Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>199606</creationdate><title>Human N‐Acetylglucosaminyltransferase III Gene is Transcribed from Multiple Promoters</title><author>Koyama, Nobuto ; 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Promoter activities of the 5′‐flanking regions of H15, H20 and exon 1 were measured in a human hepatoblastoma cell line, HuH‐6 cells by luciferase assays. The 5′‐flanking region of exon 1 was the most active, whilst that of H15 was several times less active, and that of H20 was inactive. Our study suggests that multiple promoters of the GlcNAc‐transferase III gene contribute to the complex regulation of this gene.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>8706690</pmid><doi>10.1111/j.1432-1033.1996.0853w.x</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record>
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source Wiley Online Library - AutoHoldings Journals; MEDLINE; Alma/SFX Local Collection
subjects Alternative Splicing
Base Sequence
Chromosome Mapping
DNA Primers
Gene Expression Regulation, Neoplastic
gene promoter system
genomic structure
Hepatoblastoma - genetics
Hepatoblastoma - pathology
Humans
luciferase assay
Molecular Sequence Data
N-Acetylglucosaminyltransferases - biosynthesis
N-Acetylglucosaminyltransferases - genetics
N‐acetylglucosaminyltransferase III
N‐linked oligosaccharide
Polymerase Chain Reaction - methods
Promoter Regions, Genetic
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - genetics
Sequence Analysis, DNA
Transcription, Genetic
Tumor Cells, Cultured
title Human N‐Acetylglucosaminyltransferase III Gene is Transcribed from Multiple Promoters
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