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Human androgen insensitivity mutation does not alter oligonucleotide recognition by the androgen receptor-DHT complex
Molecular and Cellular Endocrinology
|October 1, 1983
Summary
Dihydrotestosterone-receptor complexes (DHT-R) bind preferentially to specific deoxyribonucleotides like oligo-dT and -dG. Binding efficiency varies with cellular preparation and conditions, but no abnormalities were found in androgen insensitivity patients.
Area of Science:
- Molecular Endocrinology
- Genetics
- Biochemistry
Background:
- The androgen receptor (AR) mediates the biological effects of androgens.
- Understanding the DNA-binding properties of the dihydrotestosterone-receptor complex (DHT-R) is crucial for elucidating androgen action.
- Previous studies have explored AR-DNA interactions, but specific nucleotide preferences require further investigation.
Purpose of the Study:
- To investigate the binding affinity of DHT-R from human genital skin fibroblasts to various oligodeoxyribonucleotides and DNA.
- To determine the influence of different cellular preparations and experimental conditions on DHT-R binding.
- To assess potential binding abnormalities in patients with complete androgen insensitivity.
Main Methods:
- Preparation of DHT-R from total fibroblast sonicates, intact fibroblast cytosol, and nuclear extracts.
- Incubation of DHT-R with radiolabeled [3H]dihydrotestosterone.
- Binding assays using oligo-dT cellulose, DNA-cellulose, and other deoxyribonucleotides under varying salt concentrations and temperatures.
- Analysis of binding in cells from patients with receptor-negative and receptor-positive complete androgen insensitivity.
Main Results:
- DHT-R exhibited a binding preference for oligo-dT and -dG, followed by DNA, -dC, -dA, and -dI.
- Binding levels varied significantly depending on the cellular source (sonicates, cytosol, nuclear extract) and preparation conditions (temperature, salt concentration).
- No qualitative or quantitative differences in DHT-R binding to oligonucleotides or DNA were detected in cells from patients with complete androgen insensitivity.
Conclusions:
- Androgen receptor-steroid complexes demonstrate specific recognition and binding to preferred deoxyribonucleotide sequences.
- The quantitative aspects of DHT-R binding are influenced by cellular preparation methods and experimental parameters.
- Oligonucleotide and DNA binding assays, as performed, do not reveal detectable abnormalities in DHT-R from individuals with complete androgen insensitivity.