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Journal of Virology, June 2005, p. 6620-6630, Vol. 79, No. 11
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.11.6620-6630.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Selective Replication of Coronavirus Genomes That Express Nucleocapsid Protein

Barbara Schelle,1 Nadja Karl,1,{dagger} Burkhard Ludewig,2 Stuart G. Siddell,3 and Volker Thiel2*

Institute of Virology and Immunology, University of Würzburg, Würzburg, Germany,1 Research Department, Kantonal Hospital St. Gallen, St. Gallen, Switzerland,2 Department of Pathology and Microbiology, School of Medical Sciences, University of Bristol, Bristol, United Kingdom3

Received 11 November 2004/ Accepted 25 January 2005

The coronavirus nucleocapsid (N) protein is a structural protein that forms a ribonucleoprotein complex with genomic RNA. In addition to its structural role, it has been described as an RNA-binding protein that might be involved in coronavirus RNA synthesis. Here, we report a reverse genetic approach to elucidate the role of N in coronavirus replication and transcription. We found that human coronavirus 229E (HCoV-229E) vector RNAs that lack the N gene were greatly impaired in their ability to replicate, whereas the transcription of subgenomic mRNA from these vectors was easily detectable. In contrast, vector RNAs encoding a functional N protein were able to carry out both replication and transcription. Furthermore, modification of the transcription signal required for the synthesis of N protein mRNAs in the HCoV-229E genome resulted in the selective replication of genomes that are able to express the N protein. This genetic evidence leads us to conclude that at least one coronavirus structural protein, the N protein, is involved in coronavirus replication.

* Corresponding author. Mailing address: Kantonal Hospital St. Gallen, Research Department, 9007 St. Gallen, Switzerland. Phone: 41-71-4942843. Fax: 41-71-4946321. E-mail: volker.thiel{at}kssg.ch.

{dagger} Present address: Rudolf-Virchow Center for Experimental Biomedicine, 97078 Würzburg, Germany.

Journal of Virology, June 2005, p. 6620-6630, Vol. 79, No. 11
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.11.6620-6630.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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