The sup-7(st5) X gene of Caenorhabditis elegans encodes a tRNATrpUAG amber suppressor

In earlier studies, we identified in Caenorhabditis elegans two informational suppressors sup-5 III and sup-7 X and recently showed that these suppressors acted via an altered tRNA to suppress translational termination at amber (UAG) stop codons. We now show that the sup-7 (st5) suppressor is a tRNA...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1984-11, Vol.81 (21), p.6784-6788
Main Authors: S L Bolten, P Powell-Abel, D A Fischhoff, R H Waterston
Format: Article
Language:English
Subjects:
Animals
Bacteriophage lambda - genetics
Base Sequence
Caenorhabditis - genetics
DNA Restriction Enzymes
DNA, Recombinant
Mutation
Nucleic Acid Hybridization
RNA, Transfer, Amino Acyl - genetics
Suppression, Genetic
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Summary:In earlier studies, we identified in Caenorhabditis elegans two informational suppressors sup-5 III and sup-7 X and recently showed that these suppressors acted via an altered tRNA to suppress translational termination at amber (UAG) stop codons. We now show that the sup-7 (st5) suppressor is a tRNATrpUAG amber suppressor. These studies utilized a radiolabeled purified tRNA fraction to identify hybridizing genomic sequences in a phage genomic library. DNA sequence analysis of the hybridizing segment of one clone showed that the probe recognized a tRNATrpUGG sequence. The sup-7 gene was shown to be one of an 11 or 12 member tRNATrp family by Southern blot analysis, taking advantage of an Xba I restriction site induced in the anticodon sequence by the mutational event to suppressor. Sequence analysis of a recombinant lambda clone containing sup-7 gene proved that sup-7(st5) is a tRNATrpUAG. This conclusive proof of the nature of sup7(st5) will permit unambiguous interpretation in genetic applications, and the availability of the cloned sequences may allow the sup-7 gene to be used to select for the reintroduction of DNA into C. elegans.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.81.21.6784