Thumbnail
Access Restriction
Open

Author Elsasser, M. S. ♦ Eisen, L. P.
Source United States Department of Energy Office of Scientific and Technical Information
Content type Text
Language English
Subject Keyword BASIC BIOLOGICAL SCIENCES ♦ OLIGONUCLEOTIDES ♦ DNA SEQUENCING ♦ RECEPTORS ♦ BIOCHEMICAL REACTION KINETICS ♦ GLUCOCORTICOIDS ♦ LIGANDS ♦ LYMPHOCYTES ♦ MAN ♦ MUTANTS ♦ PHOSPHORUS 32 ♦ TRITIUM COMPOUNDS ♦ TUMOR CELLS ♦ ADRENAL HORMONES ♦ ANIMAL CELLS ♦ ANIMALS ♦ BETA DECAY RADIOISOTOPES ♦ BETA-MINUS DECAY RADIOISOTOPES ♦ BIOLOGICAL MATERIALS ♦ BLOOD ♦ BLOOD CELLS ♦ BODY FLUIDS ♦ CONNECTIVE TISSUE CELLS ♦ CORTICOSTEROIDS ♦ DAYS LIVING RADIOISOTOPES ♦ HORMONES ♦ HYDROGEN COMPOUNDS ♦ HYDROXY COMPOUNDS ♦ ISOTOPES ♦ KETONES ♦ KINETICS ♦ LEUKOCYTES ♦ LIGHT NUCLEI ♦ MAMMALS ♦ MATERIALS ♦ MEMBRANE PROTEINS ♦ NUCLEI ♦ NUCLEIC ACIDS ♦ ODD-ODD NUCLEI ♦ ORGANIC COMPOUNDS ♦ PHOSPHORUS ISOTOPES ♦ PREGNANES ♦ PRIMATES ♦ PROTEINS ♦ RADIOISOTOPES ♦ REACTION KINETICS ♦ SOMATIC CELLS ♦ STEROID HORMONES ♦ STEROIDS ♦ STRUCTURAL CHEMICAL ANALYSIS ♦ VERTEBRATES ♦ Biochemistry- Tracer Techniques
Abstract The stability and DNA-binding properties of activation-labile (act{sup 1}) human glucocorticoid receptors (hGRs) from the glucocorticoid-resistant mutant 3R7.6TG.4 were investigated. These receptors are able to bind reversible associating ligands with normal affinity and specificity, but become unstable during attempted activation to the DNA binding form. Affinity labeling and immunochemical analysis demonstrated that act{sup 1} receptors are not preferentially proteolyzed during attempted activation. In addition, analysis of binding to calf thymus DNA showed that after loss of ligand, act{sup 1} receptors retain the ability to bind to DNA nonspecifically. A 370 bp MMTV promoter fragment containing multiple GREs and an upstream 342 bp fragment lacking GRE sequences were used to assess the binding of act{sup 1} hGR to specific DNA sequences. Immunoadsorption of hGR-DNA complexes after incubation with {sup 32}P-end-labeled fragments showed that both normal and act{sup 1} both normal and act{sup 1} hGRs could be blocked with a synthetic oligonucleotide containing a perfect palindromic GRE, but not with an oligonucleotide in which the GRE was replaced by and ERE. Analogous results were obtained for normal and act{sup 1} hGR activated in the absence of ligand, or after incubation with the glucocorticoid antagonist RU 38486. These results suggest that sequence-specific binding of the hGR does not require the presence of bound ligand and suggest a role for the ligand in trans-activation of hormonally responsive genes.
ISSN 00062960
Educational Use Research
Learning Resource Type Article
Publisher Date 1991-11-19
Publisher Place United States
Journal Biochemistry
Volume Number 30
Issue Number 46


Open content in new tab

   Open content in new tab