Home Acta Virologica 2018 Acta Virologica Vol.62, No.1, p.68-77, 2018

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Founded: 1957
ISSN 0001-723X
E-ISSN 1336-2305

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Acta Virologica Vol.62, No.1, p.68-77, 2018

Title: A novel point mutation (L70P) inactivates poliovirus 3C protease
Author: M. UMA, S. R. HEGDE, P. P. RAO, K. NAGALEKSHMI, S. GAUTHAMI, D. KUMAR, N. R. HEGDE

Abstract: Poliovirus (PV) contains a single-stranded positive-sense RNA genome, which is translated into a single polyprotein. Viral proteases process this polyprotein to produce several individual as well as fused proteins. The major viral protease 3C cleaves at nine of the eleven cleavage sites. During the process of expressing PV 3ABC protein in Escherichia coli, we identified a 3C mutant (L70P), which lost its protease activity. This loss of function was confirmed by generating recombinant adenoviruses expressing mutant and wild-type 3C. Further, infectious PV could not be recovered from PV full-length cDNA containing the L70P mutation. However, 3C L70P mutant cDNA could complement a PV cDNA containing a 1AB deletion, producing a viable virus population containing defective complementing genomes. Structural analysis of the mutant protein indicated that the L70P mutation resulted in the loss of a hydrogen bond between two residues located within a loop between two β-sheets, potentially leading to strain on the catalytic site. We conclude that L70P inactivates 3C protease because of its close proximity to the 3C catalytic site.

Keywords: poliovirus; 3C protease; inactivating mutant; prokaryotic expression; eukaryotic expression; infectious cDNA; virus rescue
Published online: 09-Mar-2018
Year: 2018, Volume: 62, Issue: 1 Page From: 68, Page To: 77
doi:10.4149/av_2018_108


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