Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Jan;78(1):415–419. doi: 10.1073/pnas.78.1.415

Yeast tRNA precursor mutated at a splice junction is correctly processed in vivo.

D Colby, P S Leboy, C Guthrie
PMCID: PMC319064  PMID: 7017715

Abstract

Yeast mutants with decreased expression of a tRNATyr gene were obtained by selection for functional inactivation of the tyrosine-inserting ochre suppressor SUP4 and subsequent screening for production of the tRNA gene product in vivo. One mutant with reduced suppressor activity was characterized by a decreased quantity of the suppressor-specific tRNA; a precursor to this tRNA, matured at both 5' and 3' termini but still containing a 14-nucleotide intervening sequence, was present in an amount greater than 7-fold that in the parent. By RNA sequence analysis of the accumulated precursor, we have identified the mutation as an A leads to G transition at the 5' splice junction. Similar analysis of the mature tRNA produced in this mutant demonstrated that the intervening sequence was accurately excised. We conclude that the specific sequence of nucleotides at this splice junction affects the efficiency but not the fidelity of processing.

Full text

PDF
416

Images in this article

416Image
on p.416
418Image
on p.418

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abelson J. RNA processing and the intervening sequence problem. Annu Rev Biochem. 1979;48:1035–1069. doi: 10.1146/annurev.bi.48.070179.005131. [DOI] [PubMed] [Google Scholar]
  2. Borer P. N., Dengler B., Tinoco I., Jr, Uhlenbeck O. C. Stability of ribonucleic acid double-stranded helices. J Mol Biol. 1974 Jul 15;86(4):843–853. doi: 10.1016/0022-2836(74)90357-x. [DOI] [PubMed] [Google Scholar]
  3. Etcheverry T., Colby D., Guthrie C. A precursor to a minor species of yeast tRNASer contains an intervening sequence. Cell. 1979 Sep;18(1):11–26. doi: 10.1016/0092-8674(79)90349-0. [DOI] [PubMed] [Google Scholar]
  4. Goodman H. M., Olson M. V., Hall B. D. Nucleotide sequence of a mutant eukaryotic gene: the yeast tyrosine-inserting ochre suppressor SUP4-o. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5453–5457. doi: 10.1073/pnas.74.12.5453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Guthrie C., Seidman J. G., Comer M. M., Bock R. M., Schmidt F. J., Barrell B. G., McClain W. H. The biology of bacteriophage T4 transfer RNAs. Brookhaven Symp Biol. 1975 Jul;(26):106–123. [PubMed] [Google Scholar]
  6. Hopper A. K., Banks F. A yeast mutant which accumulates precursor tRNAs. Cell. 1978 Jun;14(2):211–219. doi: 10.1016/0092-8674(78)90108-3. [DOI] [PubMed] [Google Scholar]
  7. Khoury G., Gruss P., Dhar R., Lai C. J. Processing and expression of early SV40 mRNA: a role for RNA conformation in splicing. Cell. 1979 Sep;18(1):85–92. doi: 10.1016/0092-8674(79)90356-8. [DOI] [PubMed] [Google Scholar]
  8. Knapp G., Beckmann J. S., Johnson P. F., Fuhrman S. A., Abelson J. Transcription and processing of intervening sequences in yeast tRNA genes. Cell. 1978 Jun;14(2):221–236. doi: 10.1016/0092-8674(78)90109-5. [DOI] [PubMed] [Google Scholar]
  9. Laten H., Gorman J., Bock R. M. Isopentenyladenosine deficient tRNA from an antisuppressor mutant of Saccharomyces cerevisiae. Nucleic Acids Res. 1978 Nov;5(11):4329–4342. doi: 10.1093/nar/5.11.4329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lerner M. R., Steitz J. A. Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5495–5499. doi: 10.1073/pnas.76.11.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. O'Farrell P. Z., Cordell B., Valenzuela P., Rutter W. J., Goodman H. M. Structure and processing of yeast precursor tRNAs containing intervening sequences. Nature. 1978 Aug 3;274(5670):438–445. doi: 10.1038/274438a0. [DOI] [PubMed] [Google Scholar]
  12. Olson M. V., Hall B. D., Cameron J. R., Davis R. W. Cloning of the yeast tyrosine transfer RNA genes in bacteriophage lambda. J Mol Biol. 1979 Jan 25;127(3):285–295. doi: 10.1016/0022-2836(79)90330-9. [DOI] [PubMed] [Google Scholar]
  13. Rothstein R. J. A genetic fine structure analysis of the suppressor 3 locus in Saccharomyces. Genetics. 1977 Jan;85(1):55–64. doi: 10.1093/genetics/85.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Saneyoshi M., Oashi Z., Harada F., Nishimura S. Isolation and characterization of 2-methyladenosine from Escherichia coli tRNA Glu 2 , tRNA Asp 1 , tRNA His 1 and tRNA Arg . Biochim Biophys Acta. 1972 Feb 23;262(1):1–10. [PubMed] [Google Scholar]
  15. Sprinzl M., Grueter F., Spelzhaus A., Gauss D. H. Compilation of tRNA sequences. Nucleic Acids Res. 1980 Jan 11;8(1):r1–r22. [PMC free article] [PubMed] [Google Scholar]
  16. Stahl D. A., Meyhack B., Pace N. R. Recognition of local nucleotide conformation in contrast to sequence by a rRNA processing endonuclease. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5644–5648. doi: 10.1073/pnas.77.10.5644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stewart J. W., Sherman F., Jackson M., Thomas F. L., Shipman N. Demonstration of the UAA ochre codon in bakers yeast by amino-acid replacements in iso-1-cytochrome c. J Mol Biol. 1972 Jul 14;68(1):83–96. doi: 10.1016/0022-2836(72)90264-1. [DOI] [PubMed] [Google Scholar]
  18. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  19. Valenzuela P., Venegas A., Weinberg F., Bishop R., Rutter W. J. Structure of yeast phenylalanine-tRNA genes: an intervening DNA segment within the region coding for the tRNA. Proc Natl Acad Sci U S A. 1978 Jan;75(1):190–194. doi: 10.1073/pnas.75.1.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Venegas A., Quiroga M., Zaldivar J., Rutter W. J., Valenzuela P. Isolation of yeast tRNALeu genes. DNA sequence of a cloned tRNALeu3 gene. J Biol Chem. 1979 Dec 25;254(24):12306–12309. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES