Uridylylation of the genome-linked protein of poliovirus in vitro is dependent upon an endogenous RNA template.
The properties of an in vitro replication system derived from a membrane fraction (crude replication complex, CRC) of poliovirus-infected HeLa cells were examined. This system was capable of producing the nucleotidyl-proteins VPg-pU and VPg-pUpU. Due to high intrinsic phosphoesterase(s) activity and endogenous nucleoside triphosphate pools the yield of labeled product was low. Treatment of CRC with DEAE-cellulose and addition of an ATP generating system resulted in a dramatic increase in the level of nucleotidyl-proteins formed. The capacity to form VPg-pU and VPg-pUpU could be completely abolished by pretreatment of CRC with nuclease, an observation suggesting that the uridylylation of VPg is a template-dependent reaction.
Takeda
N
N
Department of Microbiology, School of Medicine, State University of New York, Stony Brook 11794.
Yang
C F
CF
Kuhn
R J
RJ
Wimmer
E
E
D005814:Genes, Viral; D006367:HeLa Cells; D006801:Humans; D017955:Poliovirus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D012367:RNA, Viral / Q000096:biosynthesis* / Q000502:physiology; D013698:Templates, Genetic; D014529:Uridine / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics; D014779:Virus Replication
D012367:RNA, Viral; D014764:Viral Proteins; D014529:Uridine