Chikungunya virus infectivity, RNA replication and non-structural polyprotein processing depend on the nsP2 protease's active site cysteine residue

Show full item record



Permalink

http://hdl.handle.net/10138/170151

Citation

Rausalu , K , Utt , A , Quirin , T , Varghese , F S , Zusinaite , E , Das , P K , Ahola , T & Merits , A 2016 , ' Chikungunya virus infectivity, RNA replication and non-structural polyprotein processing depend on the nsP2 protease's active site cysteine residue ' , Scientific Reports , vol. 6 , 37124 . https://doi.org/10.1038/srep37124

Title: Chikungunya virus infectivity, RNA replication and non-structural polyprotein processing depend on the nsP2 protease's active site cysteine residue
Author: Rausalu, Kai; Utt, Age; Quirin, Tania; Varghese, Finny S.; Zusinaite, Eva; Das, Pratyush Kumar; Ahola, Tero; Merits, Andres
Other contributor: University of Helsinki, Department of Food and Nutrition
University of Helsinki, Department of Food and Nutrition
University of Helsinki, Research Programs Unit
University of Helsinki, Department of Food and Nutrition



Date: 2016-11-15
Language: eng
Number of pages: 17
Belongs to series: Scientific Reports
ISSN: 2045-2322
DOI: https://doi.org/10.1038/srep37124
URI: http://hdl.handle.net/10138/170151
Abstract: Chikungunya virus (CHIKV), genus Alphavirus, family Togaviridae, has a positive-stand RNA genome approximately 12 kb in length. In infected cells, the genome is translated into non-structural polyprotein P1234, an inactive precursor of the viral replicase, which is activated by cleavages carried out by the non-structural protease, nsP2. We have characterized CHIKV nsP2 using both cell-free and cell-based assays. First, we show that Cys478 residue in the active site of CHIKV nsP2 is indispensable for P1234 processing. Second, the substrate requirements of CHIKV nsP2 are quite similar to those of nsP2 of related Semliki Forest virus (SFV). Third, substitution of Ser482 residue, recently reported to contribute to the protease activity of nsP2, with Ala has almost no negative effect on the protease activity of CHIKV nsP2. Fourth, Cys478 to Ala as well as Trp479 to Ala mutations in nsP2 completely abolished RNA replication in CHIKV and SFV trans-replication systems. In contrast, trans-replicases with Ser482 to Ala mutation were similar to wild type counterparts. Fifth, Cys478 to Ala as well as Trp479 to Ala mutations in nsP2 abolished the rescue of infectious virus from CHIKV RNA transcripts while Ser482 to Ala mutation had no effect. Thus, CHIKV nsP2 is a cysteine protease.
Subject: SEMLIKI-FOREST-VIRUS
SINDBIS VIRUS
TEMPORAL REGULATION
CLEAVAGE-SITE
PROTEIN NSP2
INHIBITORS
COMPLEX
IDENTIFICATION
DETERMINANTS
SPECIFICITY
3111 Biomedicine
416 Food Science
Rights:


Files in this item

Total number of downloads: Loading...

Files Size Format View
srep37124.pdf 3.205Mb PDF View/Open

This item appears in the following Collection(s)

Show full item record