The rule of six, a basic feature for efficient replication of Sendai virus defective interfering RNA.
The addition of the hepatitis delta virus genomic ribozyme to the 3' end sequence of a Sendai virus defective interfering RNA (DI-H4) allowed the reproducible and efficient replication of this RNA by the viral functions expressed from cloned genes when the DI RNA was synthesized from plasmid. Limited nucleotide additions or deletions (+7 to -7 nucleotides) in the DI RNA sequence were then made at five different sites, and the different RNA derivatives were tested for their abilities to replicate. Efficient replication was observed only when the total nucleotide number was conserved, regardless of the modifications, or when the addition of a total of 6 nucleotides was made. The replicated RNAs were shown to be properly enveloped into virus particles. It is concluded that, to form a proper template for efficient replication, the Sendai virus RNA must contain a total number of nucleotides which is a multiple of 6. This was interpreted as the need for the nucleocapsid protein to contact exactly 6 nucleotides.
Department of Genetics and Microbiology, University of Geneva Medical School, Switzerland.
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Calain
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Roux
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003673:Defective Viruses / Q000235:genetics* / Q000502:physiology; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009838:Oligodeoxyribonucleotides; D010222:Parainfluenza Virus 1, Human / Q000235:genetics* / Q000502:physiology; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D016337:RNA, Catalytic / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D015183:Restriction Mapping; D017384:Sequence Deletion; D014709:Vero Cells; D014779:Virus Replication
D009838:Oligodeoxyribonucleotides; D016337:RNA, Catalytic; D012367:RNA, Viral