Autocatalytic activity and substrate specificity of the pestivirus N-terminal protease Npro.

Gottipati, Keerthi; Acholi, Sudheer; Ruggli, Nicolas; Choi, Kyung H (2014). Autocatalytic activity and substrate specificity of the pestivirus N-terminal protease Npro. Virology, 452-453, pp. 303-309. Academic Press 10.1016/j.virol.2014.01.026

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Pestivirus N(pro) is the first protein translated in the viral polypeptide, and cleaves itself off co-translationally generating the N-terminus of the core protein. Once released, N(pro) blocks the host׳s interferon response by inducing degradation of interferon regulatory factor-3. N(pro׳)s intracellular autocatalytic activity and lack of trans-activity have hampered in vitro cleavage studies to establish its substrate specificity and the roles of individual residues. We constructed N(pro)-GFP fusion proteins that carry the authentic cleavage site and determined the autoproteolytic activities of N(pro) proteins containing substitutions at the predicted catalytic sites Glu22 and Cys69, at Arg100 that forms a salt bridge with Glu22, and at the cleavage site Cys168. Contrary to previous reports, we show that N(pro׳)s catalytic activity does not involve Glu22, which may instead be involved in protein stability. Furthermore, N(pro) does not have specificity for Cys168 at the cleavage site even though this residue is conserved throughout the pestivirus genus.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)

UniBE Contributor:

Ruggli, Nicolas

Subjects:

600 Technology > 630 Agriculture

ISSN:

0042-6822

Publisher:

Academic Press

Language:

English

Submitter:

Barbara Gautschi-Steffen

Date Deposited:

09 Apr 2015 10:41

Last Modified:

09 Apr 2015 11:02

Publisher DOI:

10.1016/j.virol.2014.01.026

PubMed ID:

24606708

Uncontrolled Keywords:

Autoprotease, Classical swine fever virus, Cysteine protease, Mass spectrometry, N-terminal protease N(pro), Pestivirus, Self-cleavage, Substrate specificity

BORIS DOI:

10.7892/boris.66672

URI:

https://boris.unibe.ch/id/eprint/66672

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