Artifacts are responsible for the translational activity of polyribosome preparations isolated from alfalfa mosaic virus-infected tobacco leaves.
We observed that polyribosome preparations isolated from alfalfa mosaic virus (A1MV)-infected tobacco leaves were contaminated with virion-derived material which could not be removed completely by sucrose gradient centrifugation or by magnesium ion precipitation. Upon incubation of polyribosome preparations with S 100 extracts from reticulocyte lysates, viral-encoded proteins were produced. Aurintricarboxylic acid (ATA), an inhibitor of initiation of translation, was used to inhibit de novo translation of the RNAS contaminating the polyribosome preparations. ATA concentrations, which did not inhibit peptide chain elongation on in vitro-produced polyribosomes, completely inhibited the translational activity of the tobacco polyribosomes. Hence the protein synthetic capacity of the tobacco polyribosome preparations is due to de novo translation of virion-derived material by vacant ribosomes present in the complementing S 100 extract. Efforts to activate the tobacco polyribosomes remain unsuccessful.
Neeleman
L
L
Department of Biochemistry, State University of Leiden, P.O. Box 9505, 2300 RA Leiden, The Netherlands.
Lindhout
P
P
van Vloten-Doting
L
L