Journal of Molecular Biology
Volume 30, Issue 2, 14 December 1967, Pages 309-322, IN17-IN18
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Initiation of protein synthesis under the direction of tobacco mosaic virus RNA in cell-free extracts of Escherichia coli

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The incorporation of amino acids under the direction of tobacco mosaic virus RNA depended upon supplementation of the cell-free Escherichia coli extracts with formyl donor, and was maximal at a magnesium ion concentration of 10 mm. The proteins made in the presence of added formyl donor were formylated. As judged by gel filtration, about 75 to 80% of the TMV product had molecular weights greater than 14,000. Up to 25% of the methionine incorporated into the TMV product was present in pronase digests as N-formylmethionine, N-formylmethionylalanine, and N-formylmethionylserine. Proteins were synthesized at half to one-quarter the rate observed in the absence of formyl donor with an optimum magnesium ion concentration between 12 and 15 mm. Under these conditions, part of the proteins lacked the formyl group, but otherwise did not appear to differ from the proteins formed in extracts supplemented with formyl donor. The product which was formed under the direction of TMV RNA was characterized by fingerprinting and gel electrophoresis. The synthesis of TMV coat protein was not detected in the presence or in the absence of added formyl donor.

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