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Encephalomyocarditis virus RNA polymerase preparations, with and without RNA helicase activity
T M Dmitrieva1, K B Norkina, V I Agol
1A. N. Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, USSR.
Abstract:
RNA template- and primer-dependent preparations of RNA polymerase were purified from encephalomyocarditis virus-infected Krebs-2 cells, using a three-step chromatographic procedure. The RNA duplex-unwinding activity of these preparations was investigated by two assays, using a partially double-stranded RNA template (encephalomyocarditis virus RNA annealed with a long segment of antisense transcript). Less purified preparations of the polymerase appeared to be able to efficiently displace, in an ATP-dependent and RNA elongation-dependent reaction, the antisense segment from the template. However, upon a more extensive purification, the unwinding activity of the RNA polymerase preparations was lost.
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