Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2001-2-22
pubmed:databankReference
pubmed:abstractText
Glutamine:fructose-6-phosphate amidotransferase (GFAT) is the first and rate-limiting enzyme of the hexosamine biosynthesis pathway, which plays an important role in glucose toxicity and cellular insulin resistance. Thus, the mechanisms by which GFAT expression is regulated under physiological and pathological conditions are of interest in connection with diabetes. In this study, we cloned the 5'-flanking region of the mouse GFAT2 gene and characterized its promoter activity. Sequence analysis revealed several putative regulatory elements Sp1, a CCAAT box, AP-1 and AP-2, but no TATA box. Transfection experiments showed that the 5'-flanking region between -2462 to +38 relative to the transcription start site of the GFAT2 gene drives transcription in NIH3T3 cells and that the fragment from -141 to -9 has the highest transcription activity. Reporter assays using deletion and mutant variants suggested that the Sp1 sites at positions -83 to -78 and -22 to -17 both play an important role in the basal promoter activity of the mouse GFAT2 gene. Electrophoretic mobility shift assay showed DNA-protein binding at both Sp1 sites. We also compared the promoter activities of mouse GFAT1 and GFAT2 in several cell lines.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0378-1119
pubmed:author
pubmed:issnType
Print
pubmed:day
31
pubmed:volume
261
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
329-36
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:11167021-3T3 Cells, pubmed-meshheading:11167021-Alkaline Phosphatase, pubmed-meshheading:11167021-Animals, pubmed-meshheading:11167021-Base Sequence, pubmed-meshheading:11167021-Binding Sites, pubmed-meshheading:11167021-Cell Line, pubmed-meshheading:11167021-Cloning, Molecular, pubmed-meshheading:11167021-DNA, pubmed-meshheading:11167021-Gene Expression Regulation, Enzymologic, pubmed-meshheading:11167021-Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing), pubmed-meshheading:11167021-Humans, pubmed-meshheading:11167021-Introns, pubmed-meshheading:11167021-Isoenzymes, pubmed-meshheading:11167021-Mice, pubmed-meshheading:11167021-Molecular Sequence Data, pubmed-meshheading:11167021-Mutagenesis, Site-Directed, pubmed-meshheading:11167021-Mutation, pubmed-meshheading:11167021-Promoter Regions, Genetic, pubmed-meshheading:11167021-Protein Binding, pubmed-meshheading:11167021-Recombinant Fusion Proteins, pubmed-meshheading:11167021-Sequence Analysis, DNA, pubmed-meshheading:11167021-Sequence Homology, Nucleic Acid, pubmed-meshheading:11167021-Sp1 Transcription Factor, pubmed-meshheading:11167021-Tumor Cells, Cultured
pubmed:year
2000
pubmed:articleTitle
Cloning and characterization of mouse glutamine:fructose-6-phosphate amidotransferase 2 gene promoter.
pubmed:affiliation
Tsukuba Research Laboratories, Eisai Co., Ltd., 5-1-3, Tokodai, Tsukuba, 300-2635, Ibaraki, Japan. k5-yamazaki@hhc.eisa.co.jp
pubmed:publicationType
Journal Article, Comparative Study