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Functional interaction between the herpes simplex-1 DNA polymerase and UL42 protein
1Department of Biochemistry, Beckman Center, Stanford University School of Medicine, California 94305-5307.
The Journal of Biological Chemistry
|July 5, 1990
Summary
The herpes simplex virus 1 UL42 protein enhances DNA polymerase processivity. This protein, along with ICP8, is crucial for complete, processive DNA replication, ensuring the generation of full-length products.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Herpes simplex virus 1 (HSV-1) genome replication requires seven viral polypeptides.
- The UL42 protein forms a 1:1 complex with HSV-1 DNA polymerase.
Purpose of the Study:
- To investigate the role of the UL42 protein in HSV-1 DNA replication.
- To characterize the processivity of HSV-1 DNA polymerase with and without UL42.
Main Methods:
- Cloning and overexpression of the HSV-1 DNA polymerase gene using a recombinant baculovirus vector.
- Purification of recombinant HSV-1 DNA polymerase.
- In vitro DNA replication assays using singly primed M13mp18 single-stranded DNA, ICP8, and purified UL42 protein.
Main Results:
- Recombinant HSV-1 DNA polymerase, in the presence of ICP8, exhibited quasi-processive polymerization, producing intermediate-length products.
- Addition of purified UL42 protein to the recombinant enzyme resulted in completely processive polymerization and full-length products.
- Processive replication by both purified native and recombinant HSV-1 DNA polymerase-UL42 complexes required saturating levels of ICP8.
Conclusions:
- The UL42 protein is essential for the processivity of HSV-1 DNA polymerase.
- ICP8 likely facilitates processivity by melting secondary structures in single-stranded DNA, enabling UL42's action.
- This study elucidates a key mechanism in HSV-1 genome replication.