Identification of crucial hydrogen-bonding residues for the interaction of herpes simplex virus DNA polymerase subunits via peptide display, mutational, and calorimetric approaches

Journal of Virology
K G BridgesDonald M Coen

Abstract

The catalytic subunit, Pol, of herpes simplex virus DNA polymerase interacts via its extreme C terminus with the processivity subunit, UL42. This interaction is critical for viral replication and thus a potential target for antiviral drug action. To investigate the Pol-binding region on UL42, we engineered UL42 mutations but also used random peptide display to identify artificial ligands of the Pol C terminus. The latter approach selected ligands with homology to residues 171 to 176 of UL42. Substitution of glutamine 171 with alanine greatly impaired binding to Pol and stimulation of long-chain DNA synthesis by Pol, identifying this residue as crucial for subunit interactions. To study these interactions quantitatively, we used isothermal titration calorimetry and wild-type and mutant forms of Pol-derived peptides and UL42. Each of three peptides corresponding to either the last 36, 27, or 18 residues of Pol bound specifically to UL42 in a 1:1 complex with a dissociation constant of 1 to 2 microM. Thus, the last 18 residues suffice for most of the binding energy, which was due mainly to a change in enthalpy. Substitutions at positions corresponding to Pol residue 1228 or 1229 or at UL42 residue 171 abolished or greatly reduced ...Continue Reading

References

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Citations

Dec 16, 2003·Journal of Molecular Biology·John C W Randell, Donald M Coen
Dec 31, 2002·Biomaterials·Yu Chi WangHsyue Jen Hsieh
Jul 29, 2008·Proceedings of the National Academy of Sciences of the United States of America·Gloria Komazin-MeredithDonald M Coen
Jan 8, 2008·The Journal of Biological Chemistry·Gloria Komazin-MeredithDonald M Coen
Aug 22, 2012·Antimicrobial Agents and Chemotherapy·Giulia MuratoreArianna Loregian
Jul 15, 2003·Journal of Virology·Arianna LoregianHoward S Marsden
Jul 19, 2002·Reviews in Medical Virology·Arianna LoregianGiorgio Palù
Aug 15, 2013·Scientific Reports·Vishnu Priyanka Reddy ChichiliJ Sivaraman
Jul 21, 2004·Molecular Cell·Brent A AppletonJames M Hogle
May 11, 2005·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·A Loregian, G Palù
Apr 2, 2003·Nature Reviews. Drug Discovery·Donald M Coen, Priscilla A Schaffer
Nov 21, 2007·Reviews in Medical Virology·Beatrice MercorelliGiorgio Palù
May 10, 2005·Journal of Cellular Physiology·Arianna Loregian, Giorgio Palù
Mar 30, 2021·Virus Research·Beata Cohan, Lori Frappier

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