Related Experiment Videos
Nonnuclear DNA binding proteins in striated muscle
J E Hagstrom1, I N Rybakova, T Staeva
1Department of Physiology, University of Wisconsin Medical School, Madison 53706, USA.
Biochemical and Molecular Medicine
|June 1, 1996
Summary
Researchers identified sarcoplasmic reticulum proteins that bind plasmid DNA in striated muscle. These DNA-binding proteins, including triadin, may play a role in muscle cell DNA uptake and expression.
Area of Science:
- Muscle physiology
- Molecular biology
- Biochemistry
Background:
- The mechanism of plasmid DNA internalization by striated muscle remains unclear.
- Non-nuclear membrane-associated DNA-binding proteins are potential mediators of DNA uptake.
- Skeletal and cardiac muscles are investigated for DNA-binding proteins.
Purpose of the Study:
- To identify and characterize DNA-binding proteins associated with the sarcoplasmic reticulum in striated muscle.
- To investigate the potential role of these proteins in plasmid DNA uptake and expression.
Main Methods:
- 22P-DNA overlay assays were employed to detect DNA-binding proteins.
- DNA-cellulose chromatography was used for protein purification and characterization.
- Subcellular fractionation and Western blotting with triadin-specific antibodies were performed.
Main Results:
- Sarcoplasmic reticulum vesicles from rabbit skeletal muscle contained 95-, 60-, and 28-kDa proteins with high-affinity double-strand DNA binding.
- These DNA-binding proteins were absent in purified sarcolemma vesicles.
- The 95- and 60-kDa proteins showed cross-reactivity with triadin-specific antibodies, suggesting they are triadin or its fragments.
Conclusions:
- Sarcoplasmic reticulum-associated proteins, including triadin, can bind plasmid DNA.
- These proteins may be involved in the transport and expression of plasmid DNA in striated muscle.
- Further research is warranted to elucidate the precise role of these proteins in gene delivery to muscle cells.