The positively charged surface of herpes simplex virus UL42 mediates DNA binding

The Journal of Biological Chemistry
Gloria Komazin-MeredithDonald M Coen

Abstract

Herpes simplex virus DNA polymerase is a heterodimer composed of UL30, a catalytic subunit, and UL42, a processivity subunit. Mutations that decrease DNA binding by UL42 decrease long chain DNA synthesis by the polymerase. The crystal structure of UL42 bound to the C terminus of UL30 revealed an extensive positively charged surface ("back face"). We tested two hypotheses, 1) the C terminus of UL30 affects DNA binding and 2) the positively charged back face mediates DNA binding. Addressing the first hypothesis, we found that the presence of a peptide corresponding to the UL30 C terminus did not result in altered binding of UL42 to DNA. Addressing the second hypothesis, previous work showed that substitution of four conserved arginine residues on the basic face with alanines resulted in decreased DNA affinity. We tested the affinities for DNA and the stimulation of long chain DNA synthesis of mutants in which the four conserved arginine residues were substituted individually or together with lysines and also a mutant in which a conserved glutamine residue was substituted with an arginine to increase positive charge on the back face. We also engineered cysteines onto this surface to permit disulfide cross-linking studies. Last, we...Continue Reading

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Citations

Jul 29, 2008·Proceedings of the National Academy of Sciences of the United States of America·Gloria Komazin-MeredithDonald M Coen
Oct 6, 2009·The Journal of Biological Chemistry·Kazutaka MurayamaShigeyuki Yokoyama
Mar 3, 2011·The Journal of Biological Chemistry·Isabella MuylaertPer Elias
Jul 12, 2011·Nucleic Acids Research·Darius Kazlauskas, Ceslovas Venclovas
Sep 7, 2012·Cold Spring Harbor Perspectives in Biology·Sandra K Weller, Donald M Coen
Nov 11, 2010·RNA·Matthew J SchellenbergAndrew M MacMillan
Aug 1, 2009·Biochemical and Biophysical Research Communications·Yu-Ching LinShih-Hsiung Wu
Aug 7, 2008·Structure·Gloria Komazin-MeredithDonald M Coen
Jun 12, 2012·Structure·Won-Ki ChoJong-Bong Lee
Nov 9, 2011·Antiviral Research·Sonia BurrelDavid Boutolleau
Mar 30, 2021·Virus Research·Beata Cohan, Lori Frappier
Dec 10, 2009·Nano Letters·Matthias BartneckJürgen Groll

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