Characterization of an equine arteritis virus replicase mutant defective in subgenomic mRNA synthesis.

Pro) is located in the nsp10 subunit and renders the EAV030F virus deficient in subgenomic mRNA synthesis. To obtain more insight into the role of nsp10 in transcription and the nature of the transcriptional defect, we have now analyzed the EAV030F mutant in considerable detail. The Ser-2429-->Pro mutation does not affect the proteolytic processing of the replicase but apparently affects the function of nsp10 in transcription. Furthermore, our study showed that EAV030F still produces subgenomic positive and negative strands, albeit at a very low level. Both subgenomic positive-strand synthesis and negative-strand synthesis are equally affected by the Ser-2429-->Pro mutation, suggesting that nsp10 plays an important role in an early step of EAV mRNA transcription. -->
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Substance Nomenclature: 0 (RNA, Messenger)
0 (RNA, Viral)
EC 2.7.7.48 (RNA-Dependent RNA Polymerase)
Entry Date(s): Date Created: 19990611 Date Completed: 19990723 Latest Revision: 20231105
Update Code: 20231215
PubMed Central ID: PMC112582
DOI: 10.1128/JVI.73.7.5274-5281.1999
PMID: 10364273
Autor: van Marle G; Department of Virology, Leiden University Medical Center, Leiden, The Netherlands., van Dinten LC, Spaan WJ, Luytjes W, Snijder EJ
Jazyk: angličtina
Zdroj: Journal of virology [J Virol] 1999 Jul; Vol. 73 (7), pp. 5274-81.
DOI: 10.1128/JVI.73.7.5274-5281.1999
Abstrakt: Equine arteritis virus (EAV) is a positive-stranded RNA virus that synthesizes a 5'- and 3'-coterminal nested set of six subgenomic mRNAs. These mRNAs all contain a common leader sequence which is derived from the 5' end of the genome. Subgenomic mRNA transcription and genome replication are directed by the viral replicase, which is expressed in the form of two polyproteins and subsequently processed into smaller nonstructural proteins (nsps). During the recent construction of an EAV infectious cDNA clone (pEAV030 [L. C. van Dinten, J. A. den Boon, A. L. M. Wassenaar, W. J. M. Spaan, and E. J. Snijder, Proc. Natl. Acad. Sci. USA 94:991-996, 1997]), a mutant cDNA clone (pEAV030F) which carries a single replicase point mutation was obtained. This substitution (Ser-2429-->Pro) is located in the nsp10 subunit and renders the EAV030F virus deficient in subgenomic mRNA synthesis. To obtain more insight into the role of nsp10 in transcription and the nature of the transcriptional defect, we have now analyzed the EAV030F mutant in considerable detail. The Ser-2429-->Pro mutation does not affect the proteolytic processing of the replicase but apparently affects the function of nsp10 in transcription. Furthermore, our study showed that EAV030F still produces subgenomic positive and negative strands, albeit at a very low level. Both subgenomic positive-strand synthesis and negative-strand synthesis are equally affected by the Ser-2429-->Pro mutation, suggesting that nsp10 plays an important role in an early step of EAV mRNA transcription.
Databáze: MEDLINE
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