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Conformational changes induced in herpes simplex virus DNA polymerase upon DNA binding
K Weisshart1, A A Kuo, G R Painter
1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Summary
Herpes simplex virus DNA polymerase undergoes significant conformational changes upon DNA binding, indicated by altered protein cleavage patterns. These changes are crucial for enzyme function and may represent a target for novel antiherpesvirus drugs.
Area of Science:
- Biochemistry
- Molecular Biology
- Virology
Background:
- Herpesvirus DNA polymerases are key targets for antiviral therapies.
- Understanding their structure-function relationship is crucial for drug development.
Purpose of the Study:
- To investigate conformational changes in herpes simplex virus DNA polymerase upon DNA binding.
- To explore the implications of these changes for enzyme activity and drug targeting.
Main Methods:
- Endogenous fluorescence quenching to detect changes in protein environment.
- Limited proteolysis with trypsin to identify conformational alterations.
Main Results:
- DNA binding reduced acrylamide quenching rate, suggesting conformational shifts.
- New trypsin cleavage sites appeared near residue 1060 and in the protein's center upon DNA/RNA binding.
- Cleavage extent correlated with ligand binding affinity.
Conclusions:
- Nucleic acid binding induces significant, localized conformational changes in herpesvirus DNA polymerase.
- These alterations likely impact substrate recognition, catalysis, and processivity.
- Targeting these conformational changes could lead to new antiherpesvirus drug strategies.