Physical and functional characterization of the double-stranded RNA binding protein encoded by the vaccinia virus E3 gene

Virology
C K Ho, S Shuman

Abstract

The vaccinia virus E3 gene encodes a 190-amino acid double-stranded (ds) RNA-binding protein that antagonizes cellular antiviral response pathways triggered by dsRNA and interferon. The physical and functional properties of the E3 protein were determined using recombinant E3 produced in bacteria and purified to homogeneity. We show by sedimentation and chemical crosslinking that E3 is a dimer in solution at high ionic strength. E3 self-associates to form higher order oligomers as ionic strength is reduced from 1 to 0.1 M NaCl. Structure probing by limited proteolysis suggests that E3 consists of amino- and carboxyl-terminal domains separated by a trypsin-sensitive bridge at residues Lys-92 and Arg-95. The carboxyl-domain of E3 contains a conserved dsRNA binding motif (dsRBM) found in many other proteins that interact with dsRNA. That the C-terminal domain per se binds to dsRNA was verified by studies of recombinant E3(100-190) purified from bacteria. The affinity of the C-terminal domain for dsRNA was comparable to that of the full-length E3 protein (KD approximately 7 to 9 nM). E3(100-190) did not bind to DNA-DNA duplexes or to DNA-RNA hybrids, suggesting that the dsRBM specifically recognizes an A-form helix. E3(100-190) is a...Continue Reading

Citations

Jul 1, 2006·Journal of Virology·Beau J FennerJimmy Kwang
Jan 5, 2002·Journal of Virology·Vincent P Smith, Antonio Alcami
May 28, 2005·Journal of Virology·Morgan Hakki, Adam P Geballe
Apr 26, 2000·Proceedings of the National Academy of Sciences of the United States of America·S McCraithS Fields
Jun 5, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yang-Gyun KimAlexander Rich
Apr 23, 2013·Virology·Craig J BierleAdam P Geballe
Sep 22, 2011·PloS One·Maude Tremblay-LétourneauMartin Bisaillon
Apr 1, 1997·Reviews in Medical Virology·M G NiemialtowskiA Schollenberger
Mar 11, 2004·The Journal of Biological Chemistry·Jana NarasimhanRonald C Wek
Sep 12, 2015·Journal of Virology·Yeu-Yang TsengWei-Li Hsu
May 29, 2003·The Journal of General Virology·Claudia González-LópezJavier Benavente
Apr 25, 1997·The Journal of Biological Chemistry·S Zhang, F Grosse
Sep 15, 1998·Methods : a Companion to Methods in Enzymology·B L JacobsT Brandt

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