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Nucleic Acids Res. 1988 April 11; 16(7): 2841–2857.
PMCID: PMC336436

A yeast tRNA(Arg) gene can act as promoter for a 5' flank deficient, non-transcribable tRNA(SUP)6 gene to produce biologically active suppressor tRNA.

Abstract

In S. cerevisiae most tRNA genes are located and expressed as single entities. The tDNA(Arg)-tDNA(Asp) pair, however, is transcribed into a dimeric precursor before being processed into two mature tRNA species. The second gene of this pair, tDNA(Asp), is totally dependent on the first gene, tDNA(Arg), and its promoter components, for homologous in vitro transcription. The second gene in the pair is now replaced by the ochre suppressor tDNA(SUP)6-o, which, by itself, cannot be transcribed because of a nonfunctional 5' flanking region. The tDNA(Arg)-tDNA(SUP)6-o was transcribed into a dimeric precursor which was processed to mature tRNA molecules as judged in vitro by electrophoretic separation, and in vivo by their ability to suppress ochre but not amber yeast mutations. Mutations in the internal promoter of the first gene decreased transcription, both in vitro and in vivo, of the second-tRNA(SUP)6-o-gene. Thus tDNA(Arg) with its 5' flanking region can act as an external promoter for other RNA polymerase III-read genes that are by themselves inactive due to impaired promoter/modulator regions.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Sentenac A. Eukaryotic RNA polymerases. CRC Crit Rev Biochem. 1985;18(1):31–90. [PubMed]
  • Sharp SJ, Schaack J, Cooley L, Burke DJ, Söll D. Structure and transcription of eukaryotic tRNA genes. CRC Crit Rev Biochem. 1985;19(2):107–144. [PubMed]
  • Newman AJ, Ogden RC, Abelson J. tRNA gene transcription in yeast: effects of specified base substitutions in the intragenic promoter. Cell. 1983 Nov;35(1):117–125. [PubMed]
  • Pearson D, Willis I, Hottinger H, Bell J, Kumar A, Leupold U, Söll D. Mutations preventing expression of sup3 tRNASer nonsense suppressors of Schizosaccharomyces pombe. Mol Cell Biol. 1985 Apr;5(4):808–815. [PMC free article] [PubMed]
  • Stewart TS, Söll D, Sharp S. Point mutations in the 5' ICR and anticodon region of a Drosophila tRNAArg gene decrease in vitro transcription. Nucleic Acids Res. 1985 Jan 25;13(2):435–447. [PMC free article] [PubMed]
  • Baker RE, Gabrielsen O, Hall BD. Effects of tRNATyr point mutations on the binding of yeast RNA polymerase III transcription factor C. J Biol Chem. 1986 Apr 25;261(12):5275–5282. [PubMed]
  • Reyes VM, Newman A, Abelson J. Mutational analysis of the coordinate expression of the yeast tRNAArg-tRNAAsp gene tandem. Mol Cell Biol. 1986 Jul;6(7):2436–2442. [PMC free article] [PubMed]
  • Larson D, Bradford-Wilcox J, Young LS, Sprague KU. A short 5' flanking region containing conserved sequences is required for silkworm alanine tRNA gene activity. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3416–3420. [PubMed]
  • Young LS, Takahashi N, Sprague KU. Upstream sequences confer distinctive transcriptional properties on genes encoding silkgland-specific tRNAAla. Proc Natl Acad Sci U S A. 1986 Jan;83(2):374–378. [PubMed]
  • Kjellin-Straby K, Engelke DR, Abelson J. Homologous in vitro transcription of linear DNA fragments containing the tRNAArg-tRNAAsp gene pair from Saccharomyces cerevisiae. DNA. 1984;3(2):167–171. [PubMed]
  • Raymond KC, Raymond GJ, Johnson JD. In vivo modulation of yeast tRNA gene expression by 5'-flanking sequences. EMBO J. 1985 Oct;4(10):2649–2656. [PubMed]
  • Dingermann T, Burke DJ, Sharp S, Schaack J, Söll D. The 5- flanking sequences of Drosophila tRNAArg genes control their in vitro transcription in a Drosophila cell extract. J Biol Chem. 1982 Dec 25;257(24):14738–14744. [PubMed]
  • Ciliberto G, Traboni C, Cortese R. Transcription of multimeric tRNA genes. Nucleic Acids Res. 1984 Jan 25;12(2):1277–1285. [PMC free article] [PubMed]
  • Allison DS, Hall BD. Effects of alterations in the 3' flanking sequence on in vivo and in vitro expression of the yeast SUP4-o tRNATyr gene. EMBO J. 1985 Oct;4(10):2657–2664. [PubMed]
  • Mao J, Schmidt O, Söll D. Dimeric transfer RNA precursors in S. pombe. Cell. 1980 Sep;21(2):509–516. [PubMed]
  • Beckmann JS, Johnson PF, Abelson J. Cloning of yeast transfer RNA genes in Escherichia coli. Science. 1977 Apr 8;196(4286):205–208. [PubMed]
  • Schmidt O, Mao J, Ogden R, Beckmann J, Sakano H, Abelson J, Söll D. Dimeric tRNA precursors in yeast. Nature. 1980 Oct 23;287(5784):750–752. [PubMed]
  • Engelke DR, Gegenheimer P, Abelson J. Nucleolytic processing of a tRNAArg-tRNAAsp dimeric precursor by a homologous component from Saccharomyces cerevisiae. J Biol Chem. 1985 Jan 25;260(2):1271–1279. [PubMed]
  • Willis I, Frendewey D, Nichols M, Hottinger-Werlen A, Schaack J, Söll D. A single base change in the intron of a serine tRNA affects the rate of RNase P cleavage in vitro and suppressor activity in vivo in Saccharomyces cerevisiae. J Biol Chem. 1986 May 5;261(13):5878–5885. [PubMed]
  • Wallace RB, Johnson PF, Tanaka S, Schöld M, Itakura K, Abelson J. Directed deletion of a yeast transfer RNA intervening sequence. Science. 1980 Sep 19;209(4463):1396–1400. [PubMed]
  • Kjellin-Straby K, Boman HG. Studies on microbial RNA, 3. Formation of submethylated sRNA in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1346–1352. [PubMed]
  • Messing J, Crea R, Seeburg PH. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. [PMC free article] [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Hinnen A, Hicks JB, Fink GR. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. [PubMed]
  • Hsiao CL, Carbon J. High-frequency transformation of yeast by plasmids containing the cloned yeast ARG4 gene. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3829–3833. [PubMed]
  • Shaw KJ, Olson MV. Effects of altered 5'-flanking sequences on the in vivo expression of a Saccharomyces cerevisiae tRNATyr gene. Mol Cell Biol. 1984 Apr;4(4):657–665. [PMC free article] [PubMed]
  • Fischhoff DA, Waterston RH, Olson MV. The yeast cloning vector YEp13 contains a tRNALeu3 gene that can mutate to an amber suppressor. Gene. 1984 Mar;27(3):239–251. [PubMed]
  • Strobel MC, Abelson J. Intron mutations affect splicing of Saccharomyces cerevisiae SUP53 precursor tRNA. Mol Cell Biol. 1986 Jul;6(7):2674–2683. [PMC free article] [PubMed]
  • Hipskind RA, Clarkson SG. 5'-flanking sequences that inhibit in vitro transcription of a xenopus laevis tRNA gene. Cell. 1983 Oct;34(3):881–890. [PubMed]
  • Frendewey D, Dingermann T, Cooley L, Söll D. Processing of precursor tRNAs in Drosophila. Processing of the 3' end involves an endonucleolytic cleavage and occurs after 5' end maturation. J Biol Chem. 1985 Jan 10;260(1):449–454. [PubMed]
  • Ratzkin B, Carbon J. Functional expression of cloned yeast DNA in Escherichia coli. Proc Natl Acad Sci U S A. 1977 Feb;74(2):487–491. [PubMed]

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