Poliovirus requires a precise 5' end for efficient positive-strand RNA synthesis.
Poliovirus infectious RNA can be synthesized in vitro using phage DNA-dependent RNA-polymerases. These synthetic transcripts contain several extra nucleotides at the 5' end, which are deleted during replication to generate authentic viral genomes. We removed those 5'-end extra nucleotides utilizing a hammerhead ribozyme to produce transcripts with accurate 5' ends. These transcripts replicate substantially more rapidly in cell culture, demonstrating no lag before replication; they also replicate more efficiently in Xenopus laevis oocytes and in in vitro translation-replication cell extracts. In both systems, an exact 5' end is necessary for synthesis of positive-strand RNA but not negative-strand RNA.
Herold
J
J
Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, California 94143-0414, USA.
Andino
R
R
D000818:Animals; D002457:Cell Extracts; D002460:Cell Line; D006801:Humans; D066298:In Vitro Techniques; D009865:Oocytes / Q000821:virology; D017955:Poliovirus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D016337:RNA, Catalytic / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis* / Q000737:chemistry; D014779:Virus Replication; D014982:Xenopus laevis
D002457:Cell Extracts; D016337:RNA, Catalytic; D012367:RNA, Viral