Linked Life Data

8626777

Source:http://linkedlifedata.com/resource/pubmed/id/8626777

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
8
pubmed:dateCreated
1996-6-21
pubmed:databankReference
pubmed:abstractText
The type II collagen gene (Col2a1) is expressed primarily in chondrocytes. Transcription of Col2a1 is mediated by cell-specific regulatory elements located within the promoter and first intron. Here, we map a minimal enhancer and identify elements that determine cartilage-specific Col2a1 expression by analyzing the activity of a series of chimeric genes consisting of rat Col2a1 first intron deletion mutants ligated to the chloramphenicol acetyltransferase reporter gene. We show that a 100-base pair (bp) segment within the first intron is the minimum size necessary for high level, cell type-specific expression of Col2a1. Sequence analysis of this 100-bp Col2a1 enhancer revealed several sequence motifs similar to motifs present within the regulatory region of the link protein gene, another cartilage gene. These motifs include an AT-rich element, a C1 motif and a C3 motif. Deletion of any of these elements reduced Col2a1 enhancer activity in chick embryo chondrocytes. We also tested enhancer-mediated activity in CFK2 cells which differentiate to a chondrogenic phenotype and begin to express type II collagen mRNA after extended culture. In stably transfected CFK2 cells, constructs containing the 100-bp enhancer were activated during the transition from prechondrogenic to chondrogenic cell populations and deletions within the enhancer strongly down-regulated activity. Chondrocyte-specific DNA-protein complexes were identified using nuclear extracts prepared from chick embryo chondrocytes and 32P-labeled oligonucleotides from these regions of the first intron. These results suggest that interaction of chondrocyte specific nuclear factors with multiple core elements from a small region within the first intron are important for cell-type specific Col2a1 enhancer activity.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
23
pubmed:volume
271
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4298-303
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:8626777-Animals, pubmed-meshheading:8626777-Base Sequence, pubmed-meshheading:8626777-Cartilage, pubmed-meshheading:8626777-Cell Nucleus, pubmed-meshheading:8626777-Chick Embryo, pubmed-meshheading:8626777-Chloramphenicol O-Acetyltransferase, pubmed-meshheading:8626777-Collagen, pubmed-meshheading:8626777-Enhancer Elements, Genetic, pubmed-meshheading:8626777-Extracellular Matrix Proteins, pubmed-meshheading:8626777-Introns, pubmed-meshheading:8626777-Molecular Sequence Data, pubmed-meshheading:8626777-Mutagenesis, pubmed-meshheading:8626777-Oligonucleotide Probes, pubmed-meshheading:8626777-Protein Biosynthesis, pubmed-meshheading:8626777-Proteins, pubmed-meshheading:8626777-Proteoglycans, pubmed-meshheading:8626777-Rats, pubmed-meshheading:8626777-Recombinant Fusion Proteins, pubmed-meshheading:8626777-Regulatory Sequences, Nucleic Acid, pubmed-meshheading:8626777-Restriction Mapping, pubmed-meshheading:8626777-Sequence Deletion, pubmed-meshheading:8626777-Sequence Homology, Nucleic Acid, pubmed-meshheading:8626777-Transfection
pubmed:year
1996
pubmed:articleTitle
Identification of a minimum enhancer sequence for the type II collagen gene reveals several core sequence motifs in common with the link protein gene.
pubmed:affiliation
Laboratory of Developmental Biology, NIDR, National Institutes of Health, Bethesda, Maryland 20892, USA.
pubmed:publicationType
Journal Article, Comparative Study