Linked Life Data

9880508

Source:http://linkedlifedata.com/resource/pubmed/id/9880508

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1999-2-11
pubmed:databankReference
pubmed:abstractText
We previously noted that the initial receptor by which murine osteoclast precursors bind matrix is the integrin alphav beta5 and that granulocyte-macrophage colony-stimulating factor (GM-CSF) decreases expression of this heterodimer by suppressing transcription of the beta5 gene. We herein report cloning of the beta5 integrin gene promoter and identification of a GM-CSF-responsive sequence. A 13-kilobase (kb) genomic fragment containing part of the beta5 gene was isolated by screening a mouse genomic library with a probe derived from the most 5'-end of a murine beta5 cDNA. A combination of primer extension and S1 nuclease studies identifies two transcriptional start sites, with the major one designated +1. A 1-kb subclone containing sequence -875 to + 110 is transcriptionally active in a murine myeloid cell line. This 1-kb fragment contains consensus binding sequences for basal (Sp1), lineage-specific (PU.1), and regulatable (signal transducer and activator of transcription) transcription factors. Reflecting our earlier findings, promoter activity is repressed in transfected myeloid cells treated with GM-CSF. Using deletion mutants, we localized a 109-base pair (bp) promoter region responsible for GM-CSF-inhibited beta5 transcription. We further identified a 19-bp sequence within the 109-bp region that binds GM-CSF-induced nuclear proteins by gel shift/competition assays. Mutation of the 19-bp sequence not only ablates its capacity to bind nuclear proteins from GM-CSF-treated cells, in vitro, but the same mutation, when introduced in the 1-kb promoter, abolishes its ability to respond to GM-CSF treatment. Northern analysis demonstrates that cycloheximide treatment abrogates the capacity of GM-CSF to decrease beta5 mRNA levels. In summary, we have identified a 19-bp cis-element mediating GM-CSF-induced down-regulation of beta5 by a mechanism requiring protein synthesis.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
274
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1366-74
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:9880508-Amino Acid Sequence, pubmed-meshheading:9880508-Animals, pubmed-meshheading:9880508-Base Sequence, pubmed-meshheading:9880508-Cloning, Molecular, pubmed-meshheading:9880508-Cycloheximide, pubmed-meshheading:9880508-Down-Regulation, pubmed-meshheading:9880508-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:9880508-Granulocyte-Macrophage Colony-Stimulating Factor, pubmed-meshheading:9880508-Humans, pubmed-meshheading:9880508-Integrin beta Chains, pubmed-meshheading:9880508-Integrins, pubmed-meshheading:9880508-Mice, pubmed-meshheading:9880508-Molecular Sequence Data, pubmed-meshheading:9880508-Point Mutation, pubmed-meshheading:9880508-Promoter Regions, Genetic, pubmed-meshheading:9880508-Protein Synthesis Inhibitors, pubmed-meshheading:9880508-Repressor Proteins, pubmed-meshheading:9880508-Transcription, Genetic
pubmed:year
1999
pubmed:articleTitle
Cloning of the murine beta5 integrin subunit promoter. Identification of a novel sequence mediating granulocyte-macrophage colony-stimulating factor-dependent repression of beta5 integrin gene transcription.
pubmed:affiliation
Departments of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
pubmed:publicationType
Journal Article