Characterization of human glucocorticoid receptor complexes formed with DNA fragments containing or lacking glucocorticoid response elements

Journal Article · Tue Mar 07 04:00:00 EST 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00431a003· OSTI ID:5649428

Tully, D B; Cidlowski, J A ^[1]

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1. Univ. of North Carolina, Chapel Hill (USA)

Sucrose density gradient shift assays were used to study the interactions of human glucocorticoid receptors (GR) with small DNA fragments either containing or lacking glucocorticoid response element (GRE) DNA consensus sequences. When crude cytoplasmic extracts containing ({sup 3}H)triamcinolone acetonide (({sup 3}H)TA) labeled GR were incubated with unlabeled DNA under conditions of DNA excess, a GRE-containing DNA fragment obtained from the 5' long terminal repeat of mouse mammary tumor virus (MMTV LTR) formed a stable 12-16S complex with activated, but not nonactivated, ({sup 3}H)TA receptor. By contrast, if the cytosols were treated with calf thymus DNA-cellulose to deplete non-GR-DNA-binding proteins prior to heat activation, a smaller 7-10S complex was formed with the MMTV LTR DNA fragment. Activated ({sup 3}H)TA receptor from DNA-cellulose pretreated cytosols also interacted with two similarly sized fragments from pBR322 DNA. Stability of the complexes formed between GR and these three DNA fragments was strongly affected by even moderate alterations in either the salt concentration or the pH of the gradient buffer. Under all conditions tested, the complex formed with the MMTV LTR DNA fragment was more stable than the complexes formed with either of the pBR322 DNA fragments. Together these observations indicate that the formation of stable complexes between activated GR and isolated DNA fragments requires the presence of GRE consensus sequences in the DNA.

• OSTI ID: 5649428
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 28:5; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADRENAL HORMONES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
CELL CONSTITUENTS
CENTRIFUGATION
CHEMICAL REACTIONS
CHEMISTRY
CORTICOSTEROIDS
CROSS-LINKING
CYTOPLASM
DAYS LIVING RADIOISOTOPES
DNA
GENE REGULATION
GLUCOCORTICOIDS
HELA CELLS
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPES
KETONES
LIGHT NUCLEI
MEMBRANE PROTEINS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
POLYMERIZATION
PREGNANES
PROTEINS
RADIOISOTOPES
RECEPTORS
SEPARATION PROCESSES
STEROIDS
TRITIUM COMPOUNDS