Linked Life Data

8702520

Source:http://linkedlifedata.com/resource/pubmed/id/8702520

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
31
pubmed:dateCreated
1996-9-24
pubmed:abstractText
We have identified sequences responsible for the expression of the human glucocorticoid receptor gene (GR gene) using a set of 5' promoter deletion mutants in HeLa, human placenta, and human breast tumor (MCF-7) cells. The chimeric gene construct -892 5'-GAAGTGACACACTTC3' -878-CAT was sufficient for high level of expression in HeLa and placenta cells in culture. Deletion of palindromic sequences decreased levels of GR expression in these cells. By oligonucleotide-affinity chromatography with the palindromic glucocorticoid receptor enhancing factor-binding element (GREFE), we have isolated from human placenta nuclear extract two novel proteins glucocorticoid receptor enhancing factors 1 and 2 (GREF1 and GREF2), with apparent molecular masses of 80 and 62 kDa, respectively. These proteins, similar to the DNA-binding autoantigen Ku are, like Ku, heterodimers of polypeptide subunits p80 and p62, immunologically related to factors binding to proximal sequence element 1 in the promoter of small nuclear RNA (PSE1) and transferrin receptor enhancing factors. Both Ku80 and Ku70 polypeptides were present in high concentrations in human placenta and HeLa cells. In MCF-7 cells, however, only a high level of p62 was detected. While cotransfection of pcDNA-Ku80 with pHGR(-892 to -878)-CAT potentiated the expression of CAT, introduction of pcDNA-Ku70 did not affect the expression of CAT in transfected MCF-7 cells. UV cross-linking analysis showed that only GREF1 contacted DNA directly. Supershift assays with monoclonal antibodies Ab 111 (Ku80) or Ab N3H10 (Ku70) showed a direct interaction of GREF1 and GREF2 heterodimers with the palindrome. Partial peptide fingerprinting of GREF1 and GREF2 using alpha-chymotrypsin and immunoblotting with Ab 111 and Ab N3H10 confirmed their identities as Ku80 and Ku70, respectively.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
2
pubmed:volume
271
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
18662-71
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:8702520-Antibodies, Monoclonal, pubmed-meshheading:8702520-Antigens, Nuclear, pubmed-meshheading:8702520-Base Sequence, pubmed-meshheading:8702520-Cell Line, pubmed-meshheading:8702520-Chimera, pubmed-meshheading:8702520-Cross-Linking Reagents, pubmed-meshheading:8702520-DNA, pubmed-meshheading:8702520-DNA Helicases, pubmed-meshheading:8702520-DNA-Binding Proteins, pubmed-meshheading:8702520-Female, pubmed-meshheading:8702520-Gene Expression, pubmed-meshheading:8702520-HeLa Cells, pubmed-meshheading:8702520-Humans, pubmed-meshheading:8702520-Immunochemistry, pubmed-meshheading:8702520-Molecular Sequence Data, pubmed-meshheading:8702520-Nuclear Proteins, pubmed-meshheading:8702520-Oligodeoxyribonucleotides, pubmed-meshheading:8702520-Peptide Mapping, pubmed-meshheading:8702520-Placenta, pubmed-meshheading:8702520-Pregnancy, pubmed-meshheading:8702520-Receptors, Glucocorticoid, pubmed-meshheading:8702520-Sequence Deletion, pubmed-meshheading:8702520-Sequence Homology, Nucleic Acid, pubmed-meshheading:8702520-Ultraviolet Rays
pubmed:year
1996
pubmed:articleTitle
Expression of human glucocorticoid receptor gene and interaction of nuclear proteins with the transcriptional control element.
pubmed:affiliation
Department of Molecular Endocrinology, Laval University Hospital Center, 2705 Boulevard Laurier, Sainte Foy, G1V 4G2, Québec, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't