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DNA-binding proteins specified by herpesvirus saimiri.
The Journal of General Virology
|December 1, 1983
Summary
Herpesvirus saimiri proteins were isolated and analyzed for DNA-binding capabilities. Several viral proteins, including structural and non-structural types, were identified as DNA binders, with similarities to Epstein-Barr virus proteins.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Herpesvirus saimiri (HVS) is a widely studied oncogenic herpesvirus.
- Understanding HVS protein interactions with DNA is crucial for deciphering viral replication and pathogenesis.
Purpose of the Study:
- To identify and characterize Herpesvirus saimiri-specific DNA-binding proteins.
- To investigate the properties of these proteins using biochemical and biophysical methods.
Main Methods:
- Nuclear protein extraction and dissociation from DNA using high salt concentrations and density gradients.
- Separation of viral proteins by differential centrifugation and Urografin gradient sedimentation.
- Analysis of DNA-binding proteins using affinity chromatography on DNA-cellulose.
- Identification of proteins by SDS-polyacrylamide gel electrophoresis and Western blotting.
Main Results:
- Several HVS-specific proteins, including structural (12K, 29K) and non-structural (51K, 76K, and others) proteins, demonstrated DNA-binding activity.
- DNA-binding proteins were eluted from DNA-cellulose columns using salt or heparin.
- The 51K phosphoprotein, 12K and 29K structural proteins, and a 100K virion polypeptide showed reproducible DNA binding.
- Binding of 130K and 110K proteins was not consistently observed after SDS-PAGE transfer.
Conclusions:
- Herpesvirus saimiri encodes multiple DNA-binding proteins essential for its lifecycle.
- These findings suggest functional similarities in DNA-binding proteins between HVS and Epstein-Barr virus.