The terminase subunits pUL56 and pUL89 of human cytomegalovirus are DNA-metabolizing proteins with toroidal structure

Nucleic Acids Research
Hanno ScheffczikE Bogner

Abstract

Herpesvirus DNA packaging involves binding and cleavage of DNA containing the specific DNA-packaging motifs. Here we report a first characterization of the terminase subunits pUL56 and pUL89 of human cytomegalovirus (HCMV). Both gene products were shown to have comparable nuclease activities in vitro. Under limiting protein concentrations the nuclease activity is enhanced by interaction of pUL56 and pUL89. High amounts of 2-bromo-5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole partially inhibited the pUL89-associated nuclease activity. It was demonstrated that pUL56 is able to bind to nucleocapsids in vivo. Electron microscopy (EM) and image analysis of purified pUL56 revealed that the molecules occurred as a distinct ring-shaped structure with a pronounced cleft. EM analysis of purified pUL89 demonstrated that this protein is also a toroidal DNA-metabolizing protein. Upon interaction of pUL56 with linearized DNA, the DNA remains uncut while the cutting event itself is mediated by pUL89. Using biochemical assays in conjunction with EM pUL56 was shown to (i) bind to DNA and (ii) associate with the capsid. In contrast to this, EM analysis implied that pUL89 is required to effect DNA cleavage. The data provide the first insights...Continue Reading

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Citations

Aug 5, 2003·Antiviral Research·Robert J Visalli, Marja van Zeijl
Sep 2, 2010·Proceedings of the National Academy of Sciences of the United States of America·Marta NadalMiquel Coll
Feb 22, 2003·Nucleic Acids Research·Brigitte ScholzElke Bogner
Jun 25, 2003·Antimicrobial Agents and Chemotherapy·Stephanie L WilliamsEarl R Kern
Sep 25, 2004·Antimicrobial Agents and Chemotherapy·David L EversJohn C Drach
Apr 24, 2004·Antimicrobial Agents and Chemotherapy·Mark R UnderwoodF Leslie Boyd
Apr 15, 2003·Journal of Virology·Philomena Ostapchuk, Patrick Hearing
Nov 26, 2005·Uirusu·Yoshito Eizuru
Apr 6, 2004·FEBS Letters·Christos G W SavvaElke Bogner
Oct 3, 2014·Chemical Reviews·Jonathan E GableCharles S Craik
May 16, 2007·Molecular Microbiology·Peixuan Guo, Tae Jin Lee
Jul 10, 2009·The Journal of General Virology·Andreas HolzenburgElke Bogner
Nov 2, 2014·The Journal of General Virology·Blair L Strang
Mar 29, 2002·Reviews in Medical Virology·Elke Bogner
Feb 19, 2004·Antiviral Chemistry & Chemotherapy·Yoshito Eizuru
Dec 6, 2017·The Journal of General Virology·Sebastian NeuberEva Maria Borst

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