Selective replication of coronavirus genomes that express nucleocapsid protein.
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.
Schelle
Barbara
B
Institute of Virology and Immunology, University of Würzburg, Germany.
Karl
Nadja
N
Ludewig
Burkhard
B
Siddell
Stuart G
SG
Thiel
Volker
V
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D028941:Coronavirus 229E, Human / Q000235:genetics* / Q000502:physiology*; D000086462:Coronavirus Nucleocapsid Proteins; D006224:Cricetinae; D004279:DNA, Viral / Q000235:genetics; D005822:Genetic Vectors; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D014779:Virus Replication / Q000235:genetics* / Q000502:physiology
D000086462:Coronavirus Nucleocapsid Proteins; D004279:DNA, Viral; D019590:Nucleocapsid Proteins; D012333:RNA, Messenger; D012367:RNA, Viral