Linked Life Data

7507230

Source:http://linkedlifedata.com/resource/pubmed/id/7507230

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1994-2-24
pubmed:databankReference
pubmed:abstractText
EndoA is a type II keratin and with EndoB (type I keratin), constitutes intermediate filaments in various simple epithelial tissues. EndoA is developmentally regulated and has an enhancer that is located at the 3'- end of the gene. This enhancer contains two single and five dual Ets binding sites. Thus far, no other promoter or enhancer has been shown to contain as many potential clustered Ets binding sites. To study the transcriptional regulation of EndoA by the ETS family proteins, we amplified the EndoA enhancer fragment from mouse genomic DNA by PCR, and cloned it into the pBLCAT2 vector upstream from the CAT reporter gene. Several pBLCAT-ENDOA clones were sequenced to verify the presence of all the ETS binding sites. Clones that did not show any point mutations in the ETS binding sites were chosen to study the transcription regulation by ETS1, ETS2 and ERGB/FLI-1 gene products. EMSA results indicated that the ETS1, ETS2 and ERGB/FLI-1 proteins bind to the enhancer sequence, and DNase I protection data demonstrated that the ETS proteins protect all seven EBS core sequences. Cotransfection of the COS cells with the pBLCAT-ENDOA construct, along with increasing amounts of different ETS expression vectors, resulted in a significant induction of CAT reporter gene expression. Previously, we have shown that the overexpression of the ETS1 gene transforms NIH3T3, and these transformed cells (7AQS2.1) produce high levels of ETS1 protein (Seth & Papas, 1990). In this report, we show that the undifferentiated P19 EC cells do not express detectable levels of ETS1; however, an elevated level of ETS1 is expressed in differentiated derivatives of these cells. We therefore used these two cell lines to examine the activity of the EndoA enhancer with the ETS1 product. Transfection of the pBLCAT-ENDOA construct alone in undifferentiated P19 EC cells results in very low CAT gene expression; however, upon differentiation with retinoic acid the level of CAT gene activity increases dramatically. Similarly, an increase in CAT expression from the same construct (pBLCAT-ENDOA) was also observed in 7AQS2.1 cells. Our results therefore indicate that the EndoA enhancer is regulated by ETS proteins via interaction with multiple ETS-binding site sequences.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0950-9232
pubmed:author
pubmed:issnType
Print
pubmed:volume
9
pubmed:geneSymbol
ETS1, ETS2, EndoA
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
469-77
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:7507230-Animals, pubmed-meshheading:7507230-Base Sequence, pubmed-meshheading:7507230-Binding Sites, pubmed-meshheading:7507230-Cell Line, pubmed-meshheading:7507230-DNA, pubmed-meshheading:7507230-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:7507230-Enhancer Elements, Genetic, pubmed-meshheading:7507230-Fibroblasts, pubmed-meshheading:7507230-Gene Expression Regulation, pubmed-meshheading:7507230-Genes, Reporter, pubmed-meshheading:7507230-Genetic Vectors, pubmed-meshheading:7507230-Insects, pubmed-meshheading:7507230-Keratins, pubmed-meshheading:7507230-Mice, pubmed-meshheading:7507230-Molecular Sequence Data, pubmed-meshheading:7507230-Polymerase Chain Reaction, pubmed-meshheading:7507230-Protein Binding, pubmed-meshheading:7507230-Proto-Oncogene Protein c-ets-1, pubmed-meshheading:7507230-Proto-Oncogene Proteins, pubmed-meshheading:7507230-Proto-Oncogene Proteins c-ets, pubmed-meshheading:7507230-Transcription Factors, pubmed-meshheading:7507230-Transcriptional Activation, pubmed-meshheading:7507230-Transfection, pubmed-meshheading:7507230-Tretinoin
pubmed:year
1994
pubmed:articleTitle
The EndoA enhancer contains multiple ETS binding site repeats and is regulated by ETS proteins.
pubmed:affiliation
Laboratory of Molecular Oncology, National Cancer Institute, Frederick, Maryland 21702-1201.
pubmed:publicationType
Journal Article