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Purification and characterization of OF-1, a host factor implicated in herpes simplex replication
R O Baker1, L B Murata, M S Dodson
1Departments of Molecular and Cellular Biology and of Biochemistry, University of Arizona, Tucson, Arizona 85721, USA.
The Journal of Biological Chemistry
|July 6, 2000
Summary
A human cellular factor, OF-1, binds to the herpes simplex virus type 1 replication origin. Its binding to Box I DNA is enhanced by the viral UL9 protein, crucial for viral DNA synthesis.
Area of Science:
- Virology
- Molecular Biology
- Cellular Biology
Background:
- Herpes simplex virus type 1 (HSV-1) replication requires specific viral and cellular factors.
- A human cellular factor, OF-1, has been previously linked to HSV-1 replication.
- OF-1 interacts with conserved regions (Boxes I, II, III) in the viral replication origin.
Purpose of the Study:
- To partially purify and characterize the human cellular factor OF-1.
- To investigate the DNA binding specificity of OF-1.
- To explore the interaction between OF-1 and the herpes simplex virus-encoded UL9 protein.
Main Methods:
- Partial purification of OF-1 from human cells.
- Mobility shift analysis to determine DNA binding specificity.
- Characterization of OF-1 protein subunits.
Main Results:
- OF-1 is a tetramer composed of 73 kDa and 90 kDa subunits; the smaller subunit possesses DNA binding activity.
- OF-1 specifically binds to both duplex and single-stranded Box I sequences, with a preference for the bottom strand.
- OF-1 binding to Box I DNA is enhanced in the presence of the viral UL9 protein.
Conclusions:
- OF-1 plays a significant role in HSV-1 replication initiation.
- The interaction between OF-1 and UL9 protein likely facilitates viral DNA synthesis.
- Understanding these interactions provides insights into the complex mechanisms of viral replication.