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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2005-1-25
pubmed:abstractText
Glycogen synthase, an enzyme involved in glycogen biosynthesis, is regulated by phosphorylation and by the allosteric ligand glucose-6-phosphate (G6P). In addition, enzyme levels can be regulated by changes in gene expression. We recently cloned a cDNA for glycogen synthase ( gsn) from Neurospora crassa, and showed that gsn transcription decreased when cells were exposed to heat shock (shifted from 30 degrees C to 45 degrees C). In order to understand the mechanisms that control gsn expression, we isolated the gene, including its 5' and 3' flanking regions, from the genome of N. crassa. An ORF of approximately 2.4 kb was identified, which is interrupted by four small introns (II-V). Intron I (482 bp) is located in the 5'UTR region. Three putative Transcription Initiation Sites (TISs) were mapped, one of which lies downstream of a canonical TATA-box sequence (5'-TGTATAAA-3'). Analysis of the 5'-flanking region revealed the presence of putative transcription factor-binding sites, including Heat Shock Elements (HSEs) and STress Responsive Elements (STREs). The possible involvement of these motifs in the negative regulation of gsn transcription was investigated using Electrophoretic Mobility Shift Assays (EMSA) with nuclear extracts of N. crassa mycelium obtained before and after heat shock, and DNA fragments encompassing HSE and STRE elements from the 5'-flanking region. While elements within the promoter region are involved in transcription under heat shock, elements in the 5'UTR intron may participate in transcription during vegetative growth. The results thus suggest that N. crassa possesses trans -acting elements that interact with the 5'-flanking region to regulate gsn transcription during heat shock and vegetative growth.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
1617-4615
pubmed:author
pubmed:issnType
Print
pubmed:volume
272
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
550-61
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:15558319-Base Sequence, pubmed-meshheading:15558319-Binding Sites, pubmed-meshheading:15558319-Cloning, Molecular, pubmed-meshheading:15558319-DNA, Complementary, pubmed-meshheading:15558319-DNA Primers, pubmed-meshheading:15558319-DNA-Binding Proteins, pubmed-meshheading:15558319-Electrophoretic Mobility Shift Assay, pubmed-meshheading:15558319-Gene Components, pubmed-meshheading:15558319-Gene Expression Regulation, Enzymologic, pubmed-meshheading:15558319-Gene Expression Regulation, Fungal, pubmed-meshheading:15558319-Genomic Library, pubmed-meshheading:15558319-Glycogen Synthase, pubmed-meshheading:15558319-Heat-Shock Response, pubmed-meshheading:15558319-Molecular Sequence Data, pubmed-meshheading:15558319-Neurospora crassa, pubmed-meshheading:15558319-Sequence Analysis, DNA
pubmed:year
2004
pubmed:articleTitle
Genomic organization of the Neurospora crassa gsn gene: possible involvement of the STRE and HSE elements in the modulation of transcription during heat shock.
pubmed:affiliation
Departamento de Bioquímica e Tecnologia Química, Instituto de Química, UNESP, 14800-900 Araraquara, SP, Brazil.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't