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Journal of Virology, May 2002, p. 4341-4349, Vol. 76, No. 9
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.9.4341-4349.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Characterization of the Protease of a Fish Retrovirus, Walleye Dermal Sarcoma Virus

Sharon K. Fodor,{dagger} and Volker M. Vogt*

Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853

Received 10 July 2001/ Accepted 4 February 2002

Three fish retroviruses infecting walleyes constitute the recently recognized genus called epsilonretrovirus. The founding member of this group, walleye dermal sarcoma virus (WDSV), induces benign skin tumors in the infected fish and replicates near 4°C. While the viral genomic sequence is known, biochemical characterization of the virus has been limited to the identification of the mature structural and envelope proteins present in virions. We undertook this study to determine the cleavage sites in the WDSV Pro and Pol proteins and to characterize the viral protease (PR) in vitro. A recombinant PR was expressed in and purified from Escherichia coli as a larger fusion with additional nucleocapsid and reverse transcriptase residues flanking the PR domain. Autocleavage produced a functional, mature PR. Autocleavage as well as cleavage of peptides and of Gag protein by the mature PR occurred at a pH optimum of 7.0, higher than that of other retroviral proteases. Analysis of the cleavage sites identified a glutamine residue in the P2 position of all WDSV sites, both in Gag and in Pol. Amino acid sequence alignments of Gag-Pro-Pol from WDSV, walleye epidermal hyperplasia virus type 1, and walleye epidermal hyperplasia virus type 2 showed the P2 glutamine to be conserved in all cleavage sites in these three viruses. Such conservation is unprecedented in other retroviruses.

* Corresponding author. Mailing address: Department of Molecular Biology and Genetics, Biotechnology Bldg., Cornell University, Ithaca, NY 14853. Phone: (607) 255-2443. Fax: (607) 255-2428. E-mail: vmv1{at}cornell.edu.

{dagger} Present address: Dept. of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109.

Journal of Virology, May 2002, p. 4341-4349, Vol. 76, No. 9
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.9.4341-4349.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Eizert, H., Bander, P., Bagossi, P., Sperka, T., Miklossy, G., Boross, P., Weber, I. T., Tozser, J. (2008). Amino Acid Preferences of Retroviral Proteases for Amino-Terminal Positions in a Type 1 Cleavage Site. J. Virol. 82: 10111-10117 [Abstract] [Full Text]  
  • Rovnak, J., Casey, R. N., Brewster, C. D., Casey, J. W., Quackenbush, S. L. (2007). Establishment of productively infected walleye dermal sarcoma explant cells. J. Gen. Virol. 88: 2583-2589 [Abstract] [Full Text]  
  • Bagossi, P., Sperka, T., Feher, A., Kadas, J., Zahuczky, G., Miklossy, G., Boross, P., Tozser, J. (2005). Amino Acid Preferences for a Critical Substrate Binding Subsite of Retroviral Proteases in Type 1 Cleavage Sites. J. Virol. 79: 4213-4218 [Abstract] [Full Text]  
  • Holzschu, D., Lapierre, L. A., Lairmore, M. D. (2003). Comparative Pathogenesis of Epsilonretroviruses. J. Virol. 77: 12385-12391 [Full Text]  


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