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Nucleic Acids Research, 1988, Vol. 16, No. 10 4569-4581
© 1988

Articles

Complementary oligodeoxynucleotide mediated inhibition of tobacco mosaic virus RNA translation in vitro

Carol Crum1, Jerry D. Johnson2, Arlen Nelson2 and Don Roth1,2,*

1Department of Plant, Soil and Insect Sciences University of Wyoming Laramie, WY 82071, USA 2Department of Molecular Biology, University of Wyoming Laramie, WY 82071, USA

* To whom correspondence should be addressed

Received September 8, 1987. Revised December 23, 1987. Accepted December 23, 1987.

Two different "antisense" oligodeoxynucleotides and their RNA analogues, each complementary to non-overlapping sequences of 51 bases near the 5' end of TMV RNA, inhibit in vitro translation of the genomic RNA in a rabbit reticulocyte lysate. Inhibition is dependent upon complementarity, concentration, and hybridization of the oligomers with TMV RNA. Inhibition is observed at molar ratios of TMV RNA to antisense oligomers as low as 1:1.5. A plateau of inhibition at which 10–25% of the control signal remains is achieved by molar ratios of TMV RNA: antisense DNA or RNA ≥1:15. The extent of inhibition is not increased by the simultaneous presence of both complementary fragments. Oligodeoxynucleotides and their RNA analogues identical to the same regions of TMV RNA have no direct effect on translation, however, they can block inhibition by the antisense fragments. Translation of BMV RNA is not affected by any of the oligodeoxynucleotides. Polyacrylamide gel electrophoresis shows translation of TMV p126 is selectively inhibited. We conclude that the observed inhibition of translation is due to direct interference with ribosome function.


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