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The herpes simplex virus processivity factor, UL42, binds DNA as a monomer.
John C W Randell1, Donald M Coen
1Committee on Virology and Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 250 Longwood Avenue, Boston, MA 02115, USA.
Journal of Molecular Biology
|December 16, 2003
Summary
Herpes simplex virus DNA polymerase processivity subunit UL42 binds DNA as a monomer. Experiments show UL42 does not form multimers on DNA, unlike similar sliding clamp proteins.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Herpes simplex virus DNA polymerase requires the UL42 protein for processivity.
- UL42 shares structural similarities with multimeric sliding clamp proteins like PCNA.
- The oligomeric state of UL42 upon DNA binding is not well understood.
Purpose of the Study:
- To determine if the UL42 protein oligomerizes when binding to DNA.
- To investigate the DNA-binding stoichiometry of UL42.
Main Methods:
- Chemical crosslinking with glutaraldehyde to detect protein-protein interactions.
- Electrophoretic mobility-shift assays (EMSA) to analyze protein:DNA complex composition.
Main Results:
- UL42 did not form intermolecular crosslinks in the presence of DNA.
- Proteins known to form multimers successfully crosslinked under the same conditions.
- EMSA results indicated that UL42 binds to DNA as a monomer.
Conclusions:
- Herpes simplex virus DNA polymerase processivity subunit UL42 functions as a monomer.
- UL42 does not oligomerize upon binding to DNA, differentiating it from other sliding clamp proteins.