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The herpes simplex virus type 1 origin-binding protein interacts specifically with the viral UL8 protein
G W McLean1, A P Abbotts, M E Parry
1Medical Research Council Virology Unit, Institute of Virology, Glasgow, U.K.
The Journal of General Virology
|October 1, 1994
Summary
Herpes simplex virus type 1 (HSV-1) helicase-primase complex proteins UL5, UL8, and UL52 were studied. Monoclonal antibody 0811 confirmed UL8
Area of Science:
- Virology
- Molecular Biology
- Protein Biochemistry
Background:
- Herpes simplex virus type 1 (HSV-1) replication relies on viral proteins.
- The UL5, UL8, and UL52 proteins form a crucial DNA helicase-primase complex.
Purpose of the Study:
- To characterize the HSV-1 helicase-primase complex.
- To investigate interactions between complex subunits and the origin-binding protein (OBP, UL9).
Main Methods:
- Purification of UL8 protein using recombinant baculovirus expression.
- Generation of monoclonal antibodies (MAbs) against UL8.
- Western blotting and immunoprecipitation assays to analyze protein interactions.
- Co-precipitation experiments using insect cells infected with recombinant viruses.
Main Results:
- MAb 0811 specifically recognized the UL8 protein.
- UL8 was shown to interact with both UL5 and UL52 proteins.
- The helicase-primase complex directly interacts with HSV-1 origin-binding protein (OBP, UL9) via the UL8 subunit.
- The DNA-binding domain of OBP (UL9) is not required for this interaction.
Conclusions:
- The UL8 subunit is a key mediator in the interaction between the HSV-1 helicase-primase complex and OBP.
- This interaction is essential for viral DNA replication initiation.