This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hausmann, Y. Articles by Rümenapf, T. Search for Related Content PubMed PubMed Citation Articles by Hausmann, Y. Articles by Rümenapf, T.

 Previous Article  |  Next Article 

Journal of Virology, May 2004, p. 5507-5512, Vol. 78, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.10.5507-5512.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Classical Swine Fever Virus Glycoprotein E^rns Is an Endoribonuclease with an Unusual Base Specificity

Yvonne Hausmann, Gleyder Roman-Sosa, Heinz-Jürgen Thiel, and Till Rümenapf^*

Institut für Virologie, Justus-Liebig-Universität, D-35392 Giessen, Germany

Received 2 October 2003/ Accepted 22 January 2004

The glycoprotein E^rns of pestiviruses is a virion-associated and -secreted RNase that is involved in virulence. The requirements at the cleavage site in heteropolymeric RNA substrates were studied for E^rns. Limited digestion of heteropolymeric RNA substrates indicated a cleavage 5' of uridine residues irrespective of the preceding nucleotide (Np/U). To further study specificity radiolabeled RNA, molecules of 45 to 56 nucleotides in length were synthesized that contained no or a single Np/U cleavage site. Cleavage was only observed in substrates containing an ApU, CpU, GpU, or UpU dinucleotide and occurred in two steps, an initial NpU-specific and a consecutive unspecific degradation. The NpU-specific cleavage was resistant to 7 M urea while the second-order cleavage was sensitive to denaturation. Kinetic analyses revealed that E^rns is a highly active endoribonuclease (k_cat/K_m = 2 x 10⁶ to 10 x 10⁶ M^–1 s^–1) with a strong affinity to NpU containing single-stranded RNA substrates (K_m = 85 to 260 nM).

---

* Corresponding author. Mailing address: Institut für Virologie, Frankfurter Str. 107, D-35392 Giessen, Germany. Phone: 49-641-99-38356. Fax: 49-641-99-38359. E-mail: Till.H.Ruemenapf{at}vetmed.uni-giessen.de.

---

Journal of Virology, May 2004, p. 5507-5512, Vol. 78, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.10.5507-5512.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Tews, B. A., Schurmann, E.-M., Meyers, G. (2009). Mutation of Cysteine 171 of Pestivirus Erns RNase Prevents Homodimer Formation and Leads to Attenuation of Classical Swine Fever Virus. J. Virol. 83: 4823-4834 [Abstract] [Full Text]  
• Tews, B. A., Meyers, G. (2007). The Pestivirus Glycoprotein Erns Is Anchored in Plane in the Membrane via an Amphipathic Helix. J. Biol. Chem. 282: 32730-32741 [Abstract] [Full Text]  
• Meyers, G., Ege, A., Fetzer, C., von Freyburg, M., Elbers, K., Carr, V., Prentice, H., Charleston, B., Schurmann, E.-M. (2007). Bovine Viral Diarrhea Virus: Prevention of Persistent Fetal Infection by a Combination of Two Mutations Affecting Erns RNase and Npro Protease. J. Virol. 81: 3327-3338 [Abstract] [Full Text]  
• Fetzer, C., Tews, B. A., Meyers, G. (2005). The Carboxy-Terminal Sequence of the Pestivirus Glycoprotein Erns Represents an Unusual Type of Membrane Anchor. J. Virol. 79: 11901-11913 [Abstract] [Full Text]  
• Lin, M., Trottier, E., Pasick, J. (2005). Antibody Responses of Pigs to Defined Erns Fragments after Infection with Classical Swine Fever Virus. CVI 12: 180-186 [Abstract] [Full Text]  
• Ivanov, K. A., Hertzig, T., Rozanov, M., Bayer, S., Thiel, V., Gorbalenya, A. E., Ziebuhr, J. (2004). Major genetic marker of nidoviruses encodes a replicative endoribonuclease. Proc. Natl. Acad. Sci. USA 101: 12694-12699 [Abstract] [Full Text]