Linked Life Data

16052479

Source:http://linkedlifedata.com/resource/pubmed/id/16052479

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2005-10-26
pubmed:abstractText
Receptor activator of nuclear factor kappa B ligand (RANKL), a potent regulator of osteoclast formation and function, is expressed by osteoblasts and bone marrow stromal cells. However, the molecular mechanism underlying RANKL expression in osteoblast/stromal cells remains largely unclear. Here, we characterized the molecular mechanism controlling RANKL basal transcription in osteoblast/stromal cells. We cloned a 1,103-bp murine RANKL promoter (from -953 to +150, relative to the transcription start site). Using a series of deletion mutants of the 1,103-bp promoter, we identified a 100-bp region (-154 to -54) mediating RANKL basal transcription in both osteoblasts and bone marrow stromal cells. Electrophoretic mobility shift assay (EMSA) using five overlapping oligonucleotides (Probes 1-5) spanning the 100-bp region showed that Probes 1 and 2 specifically bound nuclear proteins with high affinity from both cell types. Computer analysis revealed that Probes 1 and 2 contain a putative Sp1-binding site. Supershift assays with Sp1 and Sp3 antibodies confirmed that the nuclear proteins binding to Probes 1 and 2 are Sp1 and Sp3. Functionally, the mutation of the Sp1/Sp3 site in Probe 1 profoundly reduced the basal promoter activity while the mutation of the one in Probe 2 resulted in moderate reduction in the basal promoter activity. Moreover, the mutation of both sites abrogated the RANKL basal promoter activity, indicating that Sp1 and Sp3 play a key role in the RANKL basal transcription in osteoblasts and bone marrow stromal cells.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0730-2312
pubmed:author
pubmed:copyrightInfo
Copyright 2005 Wiley-Liss, Inc.
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
96
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
716-27
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:16052479-Animals, pubmed-meshheading:16052479-Base Sequence, pubmed-meshheading:16052479-Binding, Competitive, pubmed-meshheading:16052479-Binding Sites, pubmed-meshheading:16052479-Bone Marrow Cells, pubmed-meshheading:16052479-Carrier Proteins, pubmed-meshheading:16052479-Cells, Cultured, pubmed-meshheading:16052479-Consensus Sequence, pubmed-meshheading:16052479-Electrophoretic Mobility Shift Assay, pubmed-meshheading:16052479-Gene Expression Regulation, pubmed-meshheading:16052479-Membrane Glycoproteins, pubmed-meshheading:16052479-Mice, pubmed-meshheading:16052479-Molecular Sequence Data, pubmed-meshheading:16052479-Mutation, pubmed-meshheading:16052479-Nuclear Proteins, pubmed-meshheading:16052479-Oligonucleotides, pubmed-meshheading:16052479-Osteoblasts, pubmed-meshheading:16052479-Promoter Regions, Genetic, pubmed-meshheading:16052479-RANK Ligand, pubmed-meshheading:16052479-Receptor Activator of Nuclear Factor-kappa B, pubmed-meshheading:16052479-Sequence Deletion, pubmed-meshheading:16052479-Sp1 Transcription Factor, pubmed-meshheading:16052479-Sp3 Transcription Factor, pubmed-meshheading:16052479-Stromal Cells, pubmed-meshheading:16052479-Transcription, Genetic
pubmed:year
2005
pubmed:articleTitle
Sp1 and Sp3 regulate the basal transcription of receptor activator of nuclear factor kappa B ligand gene in osteoblasts and bone marrow stromal cells.
pubmed:affiliation
Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
pubmed:publicationType
Journal Article, Research Support, N.I.H., Extramural