Linked Life Data

8486650

Source:http://linkedlifedata.com/resource/pubmed/id/8486650

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
13
pubmed:dateCreated
1993-6-4
pubmed:abstractText
Transcription of the genes required for utilization of galactose in Saccharomyces cerevisiae is controlled primarily by the transcriptional activator protein GAL4. The upstream activating sequences for galactose (UASG) of most GAL genes have multiple sites to which GAL4 can bind. In this report we compare the binding properties of wild type GAL4 and derivatives of GAL4 bearing the N-terminal DNA-binding domain to multiple DNA-binding sites in vitro. To produce wild type GAL4, we constructed a recombinant baculovirus for expression in insect cells. Recombinant wild type GAL4 was found to bind efficiently to an oligonucleotide containing a near-consensus 17-mer GAL4 DNA-binding site in electrophoretic mobility shift assays. Footprinting experiments revealed that wild type GAL4 binds cooperatively to the four GAL4 DNA-binding sites of the GAL1-10 UASG; however, in contrast an N-terminal fragment of GAL4 containing only the DNA-binding/dimerization domains binds to each of these sites with slightly different affinity. With increasing concentrations of GAL4(1-147), the four sites become filled in the following order: site II, site IV, site I, and site III. In experiments with wild type GAL4, these four sites become fully occupied at approximately the same concentration of protein. In footprints of wild type GAL4 on the USAG, enhancements and protections of DNase I-sensitive cleavages are detectable between sites III and IV, indicative of formation of a loop between these distantly spaced sites. Binding of wild type GAL4 to a strong near-consensus binding site assists binding to an adjacent mutant site in both electrophoretic mobility shift and footprinting assays. GAL4(1-147) and GAL4(1-147) fused to portions of GAL4's activating region II were incapable of cooperative DNA binding in our assays. We conclude from these observations that wild type GAL4 has a cooperative DNA-binding function that is distinct from the DNA binding and dimerization or transcriptional activation functions, and likely plays and important role in precise regulation of GAL gene transcription.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
5
pubmed:volume
268
pubmed:geneSymbol
GAL, GAL1-10, GAL4
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
9629-35
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:8486650-Animals, pubmed-meshheading:8486650-Baculoviridae, pubmed-meshheading:8486650-Base Sequence, pubmed-meshheading:8486650-Binding Sites, pubmed-meshheading:8486650-Cell Line, pubmed-meshheading:8486650-DNA, Fungal, pubmed-meshheading:8486650-DNA-Binding Proteins, pubmed-meshheading:8486650-Escherichia coli, pubmed-meshheading:8486650-Fungal Proteins, pubmed-meshheading:8486650-Gene Expression Regulation, Fungal, pubmed-meshheading:8486650-Genes, Fungal, pubmed-meshheading:8486650-Genetic Vectors, pubmed-meshheading:8486650-Molecular Sequence Data, pubmed-meshheading:8486650-Moths, pubmed-meshheading:8486650-Oligodeoxyribonucleotides, pubmed-meshheading:8486650-Oligonucleotide Probes, pubmed-meshheading:8486650-Recombinant Proteins, pubmed-meshheading:8486650-Saccharomyces cerevisiae, pubmed-meshheading:8486650-Saccharomyces cerevisiae Proteins, pubmed-meshheading:8486650-Transcription, Genetic, pubmed-meshheading:8486650-Transcription Factors, pubmed-meshheading:8486650-Transfection
pubmed:year
1993
pubmed:articleTitle
Wild type GAL4 binds cooperatively to the GAL1-10 UASG in vitro.
pubmed:affiliation
Department of Biochemistry, University of British Columbia, Vancouver, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't