Purification of a terminal uridylyltransferase that acts as host factor in the in vitro poliovirus replicase reaction

Creators: Andrews, Nancy C.; Baltimore, David

Abstract

Poliovirus RNA polymerase requires a host factor to initiate RNA synthesis in vitro. The host factor was previously purified to near homogeneity from HeLa cells but was not assigned an enzymatic activity. This report describes the purification of a terminal uridylyltransferase that can act as host factor. By all criteria examined it is identical to the factor purified previously. It has the same molecular weight (68,000), chromatographic properties, and cellular localization. We present evidence that terminal uridylyltransferase can add uridine residues to the 3' poly(A) end of virion RNA and that these anneal back to the poly(A) and form a hairpin primer for polymerase.

Additional Information

© 1986 National Academy of Sciences. Contributed by David Baltimore, September 3, 1985. We thank Daniel Levin for his gift of step 6 eiF2 and double-stranded RNA-dependent protein kinase. Margaret Baron synthesized the VPg peptide used in one experiment. The Massachusetts Institute of Technology Cell Culture Center cultured HeLa cells for purification. We thank Bernard Mathey-Prevot for his advice on protein purification. N.C.A. was partially supported by a Medical Scientist Training Program grant from the National Institute of General Medical Sciences (2T 32 GM07753-06); this work was also funded by a grant from the National Institute of Allergy and Infectious Diseases (AI22346).

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