An authentic 3' noncoding region is necessary for efficient poliovirus replication.
Picornavirus RNA replication involves the specific synthesis of negative-strand intermediates followed by an accumulation of positive-strand viral RNA in the presence of a multitude of cellular mRNAs. Previously, in an effort to identify cis-acting elements required for initiation of negative-strand RNA synthesis, we deleted the entire 3' noncoding regions from human rhinovirus and poliovirus genomic RNAs. These deletion mutation transcripts displayed a severe delay in RNA accumulation following transfection of HeLa cells. Interestingly, in subsequent infection of HeLa cells, the deletion-mutant poliovirus displayed only a moderate deficiency in RNA synthesis. These data suggested that the delay in the production of cytopathic effects after transfection may have been due to an RNA replication defect overcome by the accumulation of a compensatory mutation(s) generated during initial rounds of RNA synthesis. In this study, we have sequenced the entire genome of the deletion-mutant virus and found only two nucleotide changes from the parental clone. Transfection analysis of these sequence variants revealed that the sequence changes did not provide compensatory functions for the 3' noncoding region deletion mutation replication defect. Further examination of the deletion mutant phenotype revealed that the severe replication defect following RNA transfection is due, in part, to nonviral terminal sequences present in the in vitro-derived deletion mutation transcripts. Our data suggest that poliovirus RNA harboring a complete 3' noncoding region deletion mutation is infectious (not merely quasi-infectious).
Brown
David M
DM
Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, CA 92697, USA.
Cornell
Christopher T
CT
Tran
Genevieve P
GP
Nguyen
Joseph H C
JH
Semler
Bert L
BL
D020413:3' Untranslated Regions / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D003198:Computer Simulation; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D016679:Genome, Viral; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D010641:Phenotype; D017955:Poliovirus / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology*; D014176:Protein Biosynthesis; D020871:RNA Stability; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D017384:Sequence Deletion; D014162:Transfection; D014779:Virus Replication / Q000235:genetics / Q000502:physiology
D020413:3' Untranslated Regions; D018076:DNA, Complementary; D004279:DNA, Viral; D012367:RNA, Viral