Abstract
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.
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