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Characterization of major recognition sequences for a herpes simplex virus type 1 origin-binding protein
1Department of Microbiology, State University of New York, Stony Brook 11794-8621.
Journal of Virology
|November 1, 1988
Summary
Herpes simplex virus type 1 origin-binding protein interacts with oris1 DNA. This interaction involves a specific 8-base pair sequence, suggesting a dimeric binding mechanism crucial for viral replication initiation.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Herpes simplex virus type 1 (HSV-1) replication is initiated at a specific DNA origin, oris1.
- Understanding the early protein-DNA interactions at oris1 is crucial for elucidating viral replication mechanisms.
Purpose of the Study:
- To investigate the early protein-DNA interactions at the HSV-1 origin of replication (oris1).
- To characterize the binding of cellular proteins to oris1 and identify the specific DNA sequences involved.
Main Methods:
- Mobility shift assay to detect origin-specific DNA-protein interactions.
- DNase I protection and methylation interference assays to characterize binding sites and protein-DNA contacts.
Main Results:
- Two origin-specific DNA-binding interactions were detected.
- A prominent interaction involved a protein binding to a specific 17-base pair region on both DNA strands.
- Methylation interference identified an 8-base pair binding domain (5'-GTTCGCAC-3'/3'-CAAGCGTG-5') with two overlapping inverted repeats, suggesting dimeric binding.
Conclusions:
- The identified origin-binding protein is likely related to previously defined HSV-1 origin-binding proteins.
- The specific binding of this protein to the inverted repeat sequences within oris1 is critical for initiating HSV-1 DNA replication.