Cloning and characterization of rat pancreatic β-cell/liver type glucose transporter gene: A Unique exon/intron organization

Yong Ho Ahn, Jae Woo Kim, Gil Soo Han, Byung Gwan Lee, YuSeun Kim

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Four overlapping λ genomic clones encoding rat pan- creatic β-cell/liver type glucose transporter (GLUT2) have been isolated and characterized. The gene is about 35 kb long and contains 14 exons and 13 introns. Contrary to the exon 1 of the human or mouse counterpart, the rat GLUT2 gene has three additional noncoding exons which were identified by 5′-RACE and all four were designated exon 1a, 1b, 1c, and 1d. The intron sequences bordering the splice site junctions generally follow the GT/AG rule except for one intron which begins with GC. The exon sequences determined from genomic DNA sequencing showed some differences when compared to the published rat GLUT2 cDNA. Transcription initiation site was determined by primer extension and located 661 bp upstream of the ATG translation initiation codon. Several potential binding sites for transcription factors such as C/EBP, Sp1, AP1, HNF-5, and UPE were observed and they may be responsible for the regulation of GLUT2 gene expression. The promoter region of rat GLUT2 showed little homology when compared with those of human or mouse. However, striking sequence identity (84%) was found when the adjacent intron regions flanking exon 1c were compared with the -970/-721 region of the mouse GLUT2 promoter. A series of deleted mutant constructs of the putative promoter region linked to the CAT reporter gene showed promoter activity in the primary hepatocyte culture. The region containing -452/+240 showed the highest CAT activity and fur ther deletion of the region showed gradual decrease in CAT activity.

Original languageEnglish
Pages (from-to)387-396
Number of pages10
JournalArchives of Biochemistry and Biophysics
Volume323
Issue number2
DOIs
Publication statusPublished - 1995 Nov 10

Fingerprint

Facilitative Glucose Transport Proteins
Cloning
Liver
Introns
Rats
Organism Cloning
Exons
Genes
Genetic Promoter Regions
Initiator Codon
Transcription Initiation Site
Gene Expression Regulation
DNA Sequence Analysis
Reporter Genes
Cell culture
Gene expression
Hepatocytes
Transcription Factors
Complementary DNA
Clone Cells

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

@article{b7e8ddbd8ed54f4f9312703543696690,
title = "Cloning and characterization of rat pancreatic β-cell/liver type glucose transporter gene: A Unique exon/intron organization",
abstract = "Four overlapping λ genomic clones encoding rat pan- creatic β-cell/liver type glucose transporter (GLUT2) have been isolated and characterized. The gene is about 35 kb long and contains 14 exons and 13 introns. Contrary to the exon 1 of the human or mouse counterpart, the rat GLUT2 gene has three additional noncoding exons which were identified by 5′-RACE and all four were designated exon 1a, 1b, 1c, and 1d. The intron sequences bordering the splice site junctions generally follow the GT/AG rule except for one intron which begins with GC. The exon sequences determined from genomic DNA sequencing showed some differences when compared to the published rat GLUT2 cDNA. Transcription initiation site was determined by primer extension and located 661 bp upstream of the ATG translation initiation codon. Several potential binding sites for transcription factors such as C/EBP, Sp1, AP1, HNF-5, and UPE were observed and they may be responsible for the regulation of GLUT2 gene expression. The promoter region of rat GLUT2 showed little homology when compared with those of human or mouse. However, striking sequence identity (84{\%}) was found when the adjacent intron regions flanking exon 1c were compared with the -970/-721 region of the mouse GLUT2 promoter. A series of deleted mutant constructs of the putative promoter region linked to the CAT reporter gene showed promoter activity in the primary hepatocyte culture. The region containing -452/+240 showed the highest CAT activity and fur ther deletion of the region showed gradual decrease in CAT activity.",
author = "Ahn, {Yong Ho} and Kim, {Jae Woo} and Han, {Gil Soo} and Lee, {Byung Gwan} and YuSeun Kim",
year = "1995",
month = "11",
day = "10",
doi = "10.1006/abbi.1995.0059",
language = "English",
volume = "323",
pages = "387--396",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",
number = "2",

}

Cloning and characterization of rat pancreatic β-cell/liver type glucose transporter gene : A Unique exon/intron organization. / Ahn, Yong Ho; Kim, Jae Woo; Han, Gil Soo; Lee, Byung Gwan; Kim, YuSeun.

In: Archives of Biochemistry and Biophysics, Vol. 323, No. 2, 10.11.1995, p. 387-396.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cloning and characterization of rat pancreatic β-cell/liver type glucose transporter gene

T2 - A Unique exon/intron organization

AU - Ahn, Yong Ho

AU - Kim, Jae Woo

AU - Han, Gil Soo

AU - Lee, Byung Gwan

AU - Kim, YuSeun

PY - 1995/11/10

Y1 - 1995/11/10

N2 - Four overlapping λ genomic clones encoding rat pan- creatic β-cell/liver type glucose transporter (GLUT2) have been isolated and characterized. The gene is about 35 kb long and contains 14 exons and 13 introns. Contrary to the exon 1 of the human or mouse counterpart, the rat GLUT2 gene has three additional noncoding exons which were identified by 5′-RACE and all four were designated exon 1a, 1b, 1c, and 1d. The intron sequences bordering the splice site junctions generally follow the GT/AG rule except for one intron which begins with GC. The exon sequences determined from genomic DNA sequencing showed some differences when compared to the published rat GLUT2 cDNA. Transcription initiation site was determined by primer extension and located 661 bp upstream of the ATG translation initiation codon. Several potential binding sites for transcription factors such as C/EBP, Sp1, AP1, HNF-5, and UPE were observed and they may be responsible for the regulation of GLUT2 gene expression. The promoter region of rat GLUT2 showed little homology when compared with those of human or mouse. However, striking sequence identity (84%) was found when the adjacent intron regions flanking exon 1c were compared with the -970/-721 region of the mouse GLUT2 promoter. A series of deleted mutant constructs of the putative promoter region linked to the CAT reporter gene showed promoter activity in the primary hepatocyte culture. The region containing -452/+240 showed the highest CAT activity and fur ther deletion of the region showed gradual decrease in CAT activity.

AB - Four overlapping λ genomic clones encoding rat pan- creatic β-cell/liver type glucose transporter (GLUT2) have been isolated and characterized. The gene is about 35 kb long and contains 14 exons and 13 introns. Contrary to the exon 1 of the human or mouse counterpart, the rat GLUT2 gene has three additional noncoding exons which were identified by 5′-RACE and all four were designated exon 1a, 1b, 1c, and 1d. The intron sequences bordering the splice site junctions generally follow the GT/AG rule except for one intron which begins with GC. The exon sequences determined from genomic DNA sequencing showed some differences when compared to the published rat GLUT2 cDNA. Transcription initiation site was determined by primer extension and located 661 bp upstream of the ATG translation initiation codon. Several potential binding sites for transcription factors such as C/EBP, Sp1, AP1, HNF-5, and UPE were observed and they may be responsible for the regulation of GLUT2 gene expression. The promoter region of rat GLUT2 showed little homology when compared with those of human or mouse. However, striking sequence identity (84%) was found when the adjacent intron regions flanking exon 1c were compared with the -970/-721 region of the mouse GLUT2 promoter. A series of deleted mutant constructs of the putative promoter region linked to the CAT reporter gene showed promoter activity in the primary hepatocyte culture. The region containing -452/+240 showed the highest CAT activity and fur ther deletion of the region showed gradual decrease in CAT activity.

UR - http://www.scopus.com/inward/record.url?scp=0028799359&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028799359&partnerID=8YFLogxK

U2 - 10.1006/abbi.1995.0059

DO - 10.1006/abbi.1995.0059

M3 - Article

C2 - 7487103

AN - SCOPUS:0028799359

VL - 323

SP - 387

EP - 396

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

IS - 2

ER -