Linked Life Data

11350977

Source:http://linkedlifedata.com/resource/pubmed/id/11350977

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
20
pubmed:dateCreated
2001-5-14
pubmed:abstractText
Aggrecan is a large chondroitin sulfate proteoglycan whose expression is both cell-specific and developmentally regulated. Cloning and sequencing of the 1.8-kilobase genomic 5'-flanking sequence of the chick aggrecan gene revealed the presence of potential tissue-specific control elements including a consensus sequence found in the cartilage-associated silencers, CSIIS1 and CSIIS2, that were first characterized in the type II collagen promoter sequences, as well as numerous other cis elements. Transient transfections of chick sternal chondrocytes and fibroblasts with reporter plasmids bearing progressively deleted portions of the chick aggrecan promoter and enhancer region demonstrated cell type-specific promoter activity and identified a 420-base pair region in the genomic 5-flanking region responsible for negative regulation of the aggrecan gene. In this report, three complementary methods, DNase I footprinting assays, transient transfections, and electrophoretic mobility shift assays (EMSA), provided an integral approach to better understand the regulation of the aggrecan gene. DNase I footprinting revealed that six regions of this genomic sequence bind to nuclear proteins in a tissue-specific manner. Transient transfection of reporter constructs bearing ablations of these protected sequences showed that four of the six protected sequences, which contain the sequence TCCTCC or TCCCCT, had repressor activities in transfected chick chondrocytes. Cross-competition EMSA using nuclear protein extracted from chondrocytes or fibroblasts explored the contributions of the different sequence elements in formation of DNA-protein complexes specific to cell type. This is the first parallel examination of the EMSA patterns for six functionally defined cis elements with highly similar sequences, using protein from primary cultured cells.
pubmed:grant
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
18
pubmed:volume
276
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
16894-903
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:11350977-5' Untranslated Regions, pubmed-meshheading:11350977-Aggrecans, pubmed-meshheading:11350977-Animals, pubmed-meshheading:11350977-Base Sequence, pubmed-meshheading:11350977-Cells, Cultured, pubmed-meshheading:11350977-Chickens, pubmed-meshheading:11350977-Chondrocytes, pubmed-meshheading:11350977-Cloning, Molecular, pubmed-meshheading:11350977-DNA Footprinting, pubmed-meshheading:11350977-DNA Primers, pubmed-meshheading:11350977-Enhancer Elements, Genetic, pubmed-meshheading:11350977-Extracellular Matrix Proteins, pubmed-meshheading:11350977-Gene Expression Regulation, pubmed-meshheading:11350977-Lectins, C-Type, pubmed-meshheading:11350977-Molecular Sequence Data, pubmed-meshheading:11350977-Mutagenesis, Site-Directed, pubmed-meshheading:11350977-Promoter Regions, Genetic, pubmed-meshheading:11350977-Proteoglycans, pubmed-meshheading:11350977-Recombinant Proteins
pubmed:year
2001
pubmed:articleTitle
cis elements that control the expression of chick aggrecan.
pubmed:affiliation
Department of Pediatrics, University of Chicago, Chicago, Illinois 60637, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't