Characterization of a UGA-suppressing serine tRNA from Schizosaccharomyces pombe with the help of a new in vitro assay system for eukaryotic suppressor tRNAs

J Biol Chem. 1979 Mar 10;254(5):1546-51.

Abstract

Two different allele-specific suppressor mutants of the fission yeast Schizosaccharomyces pombe produce opal (UGA) suppressor tRNAs. This was shown by the use of a new in vitro assay for eukaryotic nonsense suppression: a wheat germ extract is programmed with rabbit globin mRNAs and the readthrough products are studied. alpha-Globin is elongated upon addition of ochre (UAA) suppressor tRNAs, whereas beta-globin yields a readthrough product with opal suppressor tRNAs. This simple and very sensitive assay allowed the purification of the opal suppressor tRNA from S. pombe strain sup3-e. The pure tRNA can be aminoacylated with serine; thus, we conclude that this suppressor tRNA inserts serine in response to the UGA termination codon of pure rabbit beta-globin mRNA.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles
  • Animals
  • Ascomycota / metabolism*
  • Globins / biosynthesis
  • Mutation
  • Plants / metabolism
  • Protein Biosynthesis
  • RNA, Messenger / metabolism
  • RNA, Transfer* / genetics
  • RNA, Transfer* / metabolism
  • Rabbits
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism*
  • Serine
  • Species Specificity
  • Triticum / metabolism

Substances

  • RNA, Messenger
  • Serine
  • Globins
  • RNA, Transfer