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Herpes simplex virus DNA replication: the UL9 gene encodes an origin-binding protein
P D Olivo1, N J Nelson, M D Challberg
1Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Summary
The UL9 protein, essential for Herpes simplex virus 1 DNA replication, binds specifically to viral origins of replication. This study identifies UL9 as the key origin-binding protein, crucial for viral DNA synthesis.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Herpes simplex virus 1 (HSV-1) requires seven genes for DNA synthesis.
- Origin-dependent DNA replication is a critical process in viral life cycles.
Purpose of the Study:
- To express and characterize the UL9 protein from HSV-1.
- To determine if the UL9 protein binds to HSV-1 origins of DNA replication.
Main Methods:
- Baculovirus expression vector used to express UL9 protein in insect cells.
- Immunoassay and DNase I footprinting employed to analyze protein-DNA interactions.
Main Results:
- UL9 protein expressed in insect cells has an apparent size of 82,000 Da.
- UL9 protein specifically binds to HSV-1 origins of DNA replication (oriS and oriL).
- DNase I footprinting revealed UL9 interacts with two sites on oriS, located within a palindromic region.
Conclusions:
- The UL9 gene product is an origin-binding protein essential for HSV-1 DNA replication.
- This finding identifies UL9 as the protein responsible for previously observed origin-binding activity.