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Journal of Virology, April 2003, p. 5021-5025, Vol. 77, No. 8
0022-538X/03/$08.00+0 DOI: 10.1128/JVI.77.8.5021-5025.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
The Processing of eIF4GI by Human Rhinovirus Type 2 2Apro: Relationship to Self-Cleavage and Role of Zinc
Walter Glaser, Andrea Triendl, and Tim Skern*
Institute of Medical Biochemistry, Division of Biochemistry, University of Vienna, Vienna Bio Center, A-1030 Vienna, Austria
Received 11 November 2002/
Accepted 21 January 2003
The 2A proteinase (2Apro) of human rhinoviruses (HRVs) is a cysteine protease containing a structurally important zinc ion. In the viral polyprotein, the enzyme cleaves between the C terminus of VP1 and its own N terminus. 2Apro also processes the two isoforms of the cellular protein, eukaryotic initiation factor 4G (eIF4G). We have shown that mature HRV2 2Apro, when translated in vitro in rabbit reticulocyte lysates, efficiently cleaves eIF4GI, although the enzyme was not immediately active upon synthesis. Here, we examine the relationship between self-processing and eIF4GI cleavage. The onset of both reactions first occurred at least 10 min after initiation of protein synthesis. Furthermore, when self-processing was prevented by a specific mutation between VP1 and 2Apro, the VP1-2Apro precursor was essentially unable to cleave eIF4GI, implying that self-processing is a prerequisite for eIF4GI cleavage. 2Apro synthesized in the presence of a potent zinc chelator is inactive; however, upon addition of excess zinc, HRV2 2Apro rapidly gained activity. Finally, the presence of the zinc chelator in the culture medium can protect HeLa cells from HRV infection.
* Corresponding author. Mailing address: Institute of Medical Biochemistry, Division of Biochemistry, University of Vienna, Vienna Bio Center, Dr. Bohrgasse 9/3, A-1030 Vienna, Austria. Phone: 43 1 4277 61620. Fax: 43 1 4277 9616. E-mail:
timothy.skern{at}univie.ac.at.
Journal of Virology, April 2003, p. 5021-5025, Vol. 77, No. 8
0022-538X/03/$08.00+0 DOI: 10.1128/JVI.77.8.5021-5025.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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