Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 Jan 15;25(2):417–422. doi: 10.1093/nar/25.2.417

Sequence requirements of the bidirectional yeast TRP4 mRNA 3'-end formation signal.

C M Egli 1, K Düvel 1, N Trabesinger-Rüf 1, S Irniger 1, G H Braus 1
PMCID: PMC146438  PMID: 9016573

Abstract

The yeast TRP4 3'-end formation signal functions in both orientations in an in vivo test system. We show here that the TRP4 3'-end formation element consists of two functionally different sequence regions. One region of approximately 70 nucleotides is located in the untranslated region between the translational stop codon and the major poly(A) site. The major poly(A) site is not part of this region and can be deleted without a decrease in TRP4 3'-end formation. 5'and 3'deletions and point mutations within this region affected 3'-end formation similarly in both orientations. In the center of this region the motif TAGT is located on the antisense strand. Point mutations within this motif resulted in a drastic reduce of 3'-end formation activity in both orientations. A second region consists of the 3'-end of the TRP4 open reading frame and is required for 3'-end formation in forward orientation. A single point mutation in a TAGT motif of the TRP4 open reading frame abolished TRP4 mRNA 3'-end formation in forward orientation and had no effect on the reverse orientation.

Full Text

The Full Text of this article is available as a PDF (209.8 KB).

Selected References

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

  1. Abe A., Hiraoka Y., Fukasawa T. Signal sequence for generation of mRNA 3' end in the Saccharomyces cerevisiae GAL7 gene. EMBO J. 1990 Nov;9(11):3691–3697. doi: 10.1002/j.1460-2075.1990.tb07581.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Braus G., Furter R., Prantl F., Niederberger P., Hütter R. Arrangement of genes TRP1 and TRP3 of Saccharomyces cerevisiae strains. Arch Microbiol. 1985 Sep;142(4):383–388. doi: 10.1007/BF00491908. [DOI] [PubMed] [Google Scholar]
  3. Chen J., Moore C. Separation of factors required for cleavage and polyadenylation of yeast pre-mRNA. Mol Cell Biol. 1992 Aug;12(8):3470–3481. doi: 10.1128/mcb.12.8.3470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Egli C. M., Braus G. H. Uncoupling of mRNA 3' cleavage and polyadenylation by expression of a hammerhead ribozyme in yeast. J Biol Chem. 1994 Nov 4;269(44):27378–27383. [PubMed] [Google Scholar]
  5. Egli C. M., Springer C., Braus G. H. A complex unidirectional signal element mediates GCN4 mRNA 3' end formation in Saccharomyces cerevisiae. Mol Cell Biol. 1995 May;15(5):2466–2473. doi: 10.1128/mcb.15.5.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  7. Furter R., Paravicini G., Aebi M., Braus G., Prantl F., Niederberger P., Hütter R. The TRP4 gene of Saccharomyces cerevisiae: isolation and structural analysis. Nucleic Acids Res. 1986 Aug 26;14(16):6357–6373. doi: 10.1093/nar/14.16.6357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giebel L. B., Spritz R. A. Site-directed mutagenesis using a double-stranded DNA fragment as a PCR primer. Nucleic Acids Res. 1990 Aug 25;18(16):4947–4947. doi: 10.1093/nar/18.16.4947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Guo Z., Russo P., Yun D. F., Butler J. S., Sherman F. Redundant 3' end-forming signals for the yeast CYC1 mRNA. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4211–4214. doi: 10.1073/pnas.92.10.4211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Guo Z., Sherman F. 3'-end-forming signals of yeast mRNA. Mol Cell Biol. 1995 Nov;15(11):5983–5990. doi: 10.1128/mcb.15.11.5983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heidmann S., Schindewolf C., Stumpf G., Domdey H. Flexibility and interchangeability of polyadenylation signals in Saccharomyces cerevisiae. Mol Cell Biol. 1994 Jul;14(7):4633–4642. doi: 10.1128/mcb.14.7.4633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Henikoff S., Cohen E. H. Sequences responsible for transcription termination on a gene segment in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1515–1520. doi: 10.1128/mcb.4.8.1515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hyman L. E., Seiler S. H., Whoriskey J., Moore C. L. Point mutations upstream of the yeast ADH2 poly(A) site significantly reduce the efficiency of 3'-end formation. Mol Cell Biol. 1991 Apr;11(4):2004–2012. doi: 10.1128/mcb.11.4.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Irniger S., Braus G. H. Saturation mutagenesis of a polyadenylation signal reveals a hexanucleotide element essential for mRNA 3' end formation in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):257–261. doi: 10.1073/pnas.91.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Irniger S., Egli C. M., Braus G. H. Different classes of polyadenylation sites in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1991 Jun;11(6):3060–3069. doi: 10.1128/mcb.11.6.3060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Irniger S., Sanfaçon H., Egli C. M., Braus G. H. Different sequence elements are required for function of the cauliflower mosaic virus polyadenylation site in Saccharomyces cerevisiae compared with in plants. Mol Cell Biol. 1992 May;12(5):2322–2330. doi: 10.1128/mcb.12.5.2322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miozzari G., Niederberger P., Hütter R. Tryptophan biosynthesis in Saccharomyces cerevisiae: control of the flux through the pathway. J Bacteriol. 1978 Apr;134(1):48–59. doi: 10.1128/jb.134.1.48-59.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Oste C. Polymerase chain reaction. Biotechniques. 1988 Feb;6(2):162–167. [PubMed] [Google Scholar]
  20. Russo P., Li W. Z., Guo Z., Sherman F. Signals that produce 3' termini in CYC1 mRNA of the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1993 Dec;13(12):7836–7849. doi: 10.1128/mcb.13.12.7836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Russo P., Li W. Z., Hampsey D. M., Zaret K. S., Sherman F. Distinct cis-acting signals enhance 3' endpoint formation of CYC1 mRNA in the yeast Saccharomyces cerevisiae. EMBO J. 1991 Mar;10(3):563–571. doi: 10.1002/j.1460-2075.1991.tb07983.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sadhale P. P., Platt T. Unusual aspects of in vitro RNA processing in the 3' regions of the GAL1, GAL7, and GAL10 genes in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Oct;12(10):4262–4270. doi: 10.1128/mcb.12.10.4262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  25. Wahle E., Keller W. The biochemistry of 3'-end cleavage and polyadenylation of messenger RNA precursors. Annu Rev Biochem. 1992;61:419–440. doi: 10.1146/annurev.bi.61.070192.002223. [DOI] [PubMed] [Google Scholar]
  26. Yu K., Elder R. T. Some of the signals for 3'-end formation in transcription of the Saccharomyces cerevisiae Ty-D15 element are immediately downstream of the initiation site. Mol Cell Biol. 1989 Jun;9(6):2431–2444. doi: 10.1128/mcb.9.6.2431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES