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Mutations in a herpes simplex virus type 1 origin that inhibit interaction with origin-binding protein also inhibit
T R Hernandez1, R E Dutch, I R Lehman
1Department of Biochemistry, Stanford University School of Medicine, California 94305-5307.
Journal of Virology
|March 1, 1991
Summary
Herpes simplex virus type 1 DNA replication relies on the origin-binding protein binding to specific sites within the OriS replication origin. Mutations disrupting this interaction significantly impair viral DNA replication.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Herpes simplex virus type 1 (HSV-1) possesses three replication origins: OriL and a diploid OriS.
- The UL9 gene product, an origin-binding protein, is crucial for viral DNA replication initiation.
- Specific DNA sequences within OriS are recognized by the origin-binding protein.
Purpose of the Study:
- To investigate the role of specific binding sites (boxes I, II, and III) within the OriS replication origin of HSV-1.
- To determine the relationship between origin-binding protein interaction with OriS and the efficiency of HSV-1 DNA replication.
Main Methods:
- Transient replication assays were used to measure OriS-directed plasmid replication.
- Site-directed mutagenesis was employed to alter sequences within the OriS binding sites.
- Correlation analysis was performed between protein-binding ability and replication efficiency.
Main Results:
- Mutations in boxes I, II, and III of OriS significantly reduced OriS-directed plasmid replication.
- Reduced replication efficiency in mutants correlated with decreased origin-binding protein binding.
- Multiple mutations suggested multiple binding sites for the origin-binding protein within OriS.
Conclusions:
- The origin-binding protein's interaction with OriS sequences is essential for HSV-1 DNA replication.
- Proper binding of the origin-binding protein to multiple sites within OriS is critical for efficient viral DNA replication.