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Affinity purification of Alu-DNA-repeat-binding proteins from human somatic cells
D V Luk'yanov1, G F Reshetnikova, O I Podgornaya
1Institute of Cytology, Russian Academy of Sciences, St. Petersburg, 194064, Russia.
Biochemistry. Biokhimiia
|February 13, 1999
Summary
Researchers identified a 66-kD Alu-DNA-binding protein in human cells. This protein shares characteristics with a similar protein found in sperm, suggesting a potential homology between germ and somatic cell proteins.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Alu DNA repeats are abundant in the human genome.
- Specific proteins are known to bind Alu sequences, but their roles in somatic cells are not fully understood.
Purpose of the Study:
- To identify and characterize proteins that bind to Alu DNA repeats in human somatic cell nucleoplasm.
- To compare the identified somatic cell protein with known Alu-binding proteins from germ cells.
Main Methods:
- Gel shift assay to study DNA-protein interactions.
- Southwestern blotting to identify protein molecular weights.
- Affinity purification using DNA-coupled carriers for protein isolation.
Main Results:
- A 66-kD protein binding to Alu DNA repeats was isolated from human somatic cell nucleoplasm.
- A 60-kD protein was found to copurify with the 66-kD protein, indicating potential protein-protein interactions.
- The binding site was localized to a specific region within the RNA polymerase III promoter, similar to that of a protein from human sperm.
Conclusions:
- The identified 66-kD somatic cell protein exhibits properties similar to the Alu-binding protein from human spermatozoids.
- The findings suggest a potential homology between Alu-binding proteins in germ and somatic cells.
- Further research is warranted to elucidate the functional relationship and evolutionary significance of these proteins.