Related Experiment Videos
The herpes simplex virus 1 origin binding protein: a DNA helicase
R C Bruckner1, J J Crute, M S Dodson
1Department of Biochemistry, Beckman Center, Stanford University, California 94305-5307.
The Journal of Biological Chemistry
|February 5, 1991
Summary
Researchers purified a herpes simplex virus type 1 (HSV-1) origin binding protein (UL9). This protein exhibits DNA helicase and nucleoside triphosphatase activities, crucial for viral DNA replication.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Herpes simplex virus type 1 (HSV-1) requires specific proteins for viral DNA replication.
- The UL9 gene product is known to bind to the HSV-1 origin of replication.
Purpose of the Study:
- To overexpress and purify the recombinant HSV-1 origin binding protein (UL9).
- To characterize the enzymatic activities of the purified recombinant UL9 protein.
Main Methods:
- Overexpression of the HSV-1 UL9 gene in mammalian cells.
- Purification of the recombinant UL9 protein to near homogeneity.
- Assays to determine DNA binding, nucleoside triphosphatase, and DNA helicase activities.
Main Results:
- The recombinant UL9 protein was successfully purified.
- The purified protein demonstrated DNA-dependent nucleoside 5'-triphosphatase and DNA helicase activities, in addition to origin binding.
- Nucleotide hydrolysis was significantly influenced by the DNA cofactor's structure and sequence.
- The properties of the recombinant protein matched those of the native protein found in infected cells.
Conclusions:
- The recombinant UL9 protein serves as a valuable tool for studying HSV-1 DNA replication.
- UL9 possesses multiple enzymatic activities essential for viral DNA replication.
- The DNA cofactor plays a critical role in regulating UL9's nucleoside triphosphatase activity.