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Properties of Single-stranded RNA Synthesized by a Crude RNA Polymerase Fraction from Barley Leaves Infected with Brome Mosaic Virus

Faculté des Sciences agronomiques 5800 Gembloux, Belgium

The RNA polymerase activity of a cell-free particulate fraction of barley leaves infected with brome mosaic virus was investigated in the presence of actinomycin D and EDTA. The RNA products were fractionated according to their solubility in 2 M-LiCl. Both LiCl-soluble and LiCl-insoluble fractions of the RNA synthesized during a 2 min. pulse of [³H]-UTP were largely resistant to RNase in high salt concentration; the radioactive RNA of the insoluble fraction had sedimentation properties expected for replicative intermediates. After a 2 min. chase, LiCl-insoluble radioactivity was essentially RNase-sensitive in high salt, and sedimented in sucrose gradients in association with all three components of brome mosaic virus RNA, provided protective exogenous RNA was added throughout the experimental procedure. The overall results suggest that the in vitro synthesis of the RNAs found in virus particles occurs by a continuous process resulting possibly from recycling activity of the polymerase.

Received 12 January 1972; accepted 18 February 1972.