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Nucleic Acids Research, 1995, Vol. 23, No. 22 4591-4597
© 1995

Articles

Cysteine tRNAs of plant origin as novel UGA suppressors

Carsten Urban and Hildburg Beier*

Institut fur Biochemie, Bayerische Julius-Maximilians-Universitàt Biozentrum, Am Hubland, D-97074 Wurzburg, Germany

*To whom correspondence should be addressed

Received September 6, 1995. Accepted October 16, 1995.

We have isolated and sequenced chloroplast (chl) and cytoplasmic (cyt) cysteine tRNAs from Nicotiana rustics. Both tRNAs carry a GCA antlcodon but beyond that differ considerably in their nucleotide sequences. One obvious distinction resides in the presence of N6-isopentenyladenoslne (i6A) and 1-methylguanosine (m1G) at position 37 in chl and cyt tRNACys respectively. In order to study the potential suppressor activity of tRNAsCys we used in vitro synthesized zein mRNA transcripts in which an internal UGA stop codon had been placed in either the tobacco rattle virus (TRV) or tobacco mosaic virus (TMV)specific codon context. In vitro translation was carried out in a messenger- and tRNA-dependent wheat germ extract. Both tRNACys isoacceptors stimulate read-through over the UGA stop codon, however, chl tRNA is more efficient than the cytoplasmic counterpart. The UGA in the two viral codon contexts is suppressed to about the same extent by either of the two tRNAs, whereas UGA in the (i-globin context is not recognized at all. The Interaction of tRNAGCACys with UGA requires an unconventional G: A base pair in the wobble position, as postulated earlier for plant tRNAG{psi}ACys misreading the UAA stop codon. This is the first case that a cysteineaccepting tRNA has been characterized as a natural UGA suppressor.


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