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J Gen Virol 76 (1995), 2169-2180; DOI 10.1099/0022-1317-76-9-2169
© 1995 Society for General Microbiology

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Analysis of cross reactivity of retrovirus proteases using a vaccinia virus-T7 RNA polymerase-based expression system

B. G. Mattias Luukkonen{dagger}, Wei Tan, Eva Maria Fenyö and Stefan Schwartz*

Microbiology and Tumorbiology Center, Karolinska Institute, PO Box 280, S-171 77 Stockholm, Sweden

We have used the vaccinia virus-T7 RNA polymerase-based expression system for studies on the activity of proteases from various retroviruses on homologous and heterologous Gag polyproteins in eukaryotic cells. Proteases from human immunodeficiency virus (HIV) types 1 and 2, equine infectious anaemia virus, human T cell leukaemia virus type 1 and human spumavirus were produced and were shown to cleave their cognate Gag substrates produced in trans. Analysis of cross reactivity revealed that lentivirus proteases cleaved only lentivirus Gag proteins and oncovirus proteases acted primarily on oncovirus Gag proteins. The HIV-2 protease cleaved the HIV-1 Gag precursor almost as efficiently as HIV-1 protease. Expression of the 5' end of the human spumavirus pol gene revealed that it encodes a functional protease that acts specifically on the human spumavirus Gag polyprotein. This assay will allow further investigation on the activity and specificity of retrovirus proteases in eukaryotic cells.

* Author for correspondence. Fax +46 8 331 399. e-mail stefan.schwartz@mtc.ki.se

{dagger} Present address: European Molecular Biology Laboratory, Meyerhofstrasse 1, 69012 Heidelberg, Germany.

Received 5 January 1995; accepted 3 May 1995.


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