Translation of encephalomyocarditis virus RNA in reticulocyte lysates: kinetic analysis of the formation of virion proteins and a protein required for processing

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Abstract
In cell-free extracts derived from rabbit reticulocytes, encephalomyocarditis RNA can be translated completely and the products can be porcessed extensively to give encephalomyocarditis virion proteins and several nonvirion proteins, including a genome-coded protein required for processing. The lateer is probably a protease. Translation is very efficient. Under typical conditions, each EMC RNA is translated about 8 times during 3 h. Kinetic analyses (time-course experiments, pulse-chase experiments and pulse-stop experiments) were used to determine the time of appearance of major products and these times were correlated with map positions. The gene for the putative protease is located near the middle of the genome downstream from the virion protein genes. Ribosomes can travel the length of encephalomyocarditis RNA within 30 min but there is a delay in their progress along the RNA at some point soon after they traverse the region coding for virion protein precursors. This delay results in the accumulation of precursors for a period of about 10 min before the putative protease is made and virion proteins (.epsilon., .alpha. and .gamma.) are released by proteolysis.