The C-terminal silencing domain of Rap1p is essential for the repression of ribosomal protein genes in response to a defect in the secretory pathway (original) (raw)

Abstract

We have previously shown that a functional secretory pathway is essential for continued ribosome synthesis in Saccharomyces cerevisiae. When a temperature-sensitive mutant defective in the secretory pathway is transferred to the non-permissive temperature, transcription of both rRNA genes and ribosomal protein genes is nearly abolished. In order to define the cis -acting element(s) of ribosomal protein genes sensitive to a defect in the secretory pathway, we have constructed a series of fusion genes containing the CYH2 promoter region, with various deletions, fused to lacZ. Each fusion gene for which transcription is detected is subject to the repression. Rap1p is the transcriptional activator for most ribosomal protein genes, as well as having an important role in silencing in the vicinity of telomeres and at the silent mating-type loci. To assess its role in the repression of transcription by the defect in the secretory pathway, we have introduced rap1 mutations. The replacement of wild-type Rap1p by Rap1p truncated at the C-terminal region caused substantial attenuation of the repression. Furthermore, we have demonstrated that the Rap1p-truncation affects the repression of TCM1 , encoding ribosomal protein L3, which has no Rap1p-binding site in its upstream regulatory region. These results suggest that the repression of transcription of ribosomal protein genes by a secretory defect is mediated through Rap1p, but does not require a Rap1p-binding site within the UAS.

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Selected References

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  1. Buck S. W., Shore D. Action of a RAP1 carboxy-terminal silencing domain reveals an underlying competition between HMR and telomeres in yeast. Genes Dev. 1995 Feb 1;9(3):370–384. doi: 10.1101/gad.9.3.370. [DOI] [PubMed] [Google Scholar]
  2. Chan C. S., Tye B. K. Organization of DNA sequences and replication origins at yeast telomeres. Cell. 1983 Jun;33(2):563–573. doi: 10.1016/0092-8674(83)90437-3. [DOI] [PubMed] [Google Scholar]
  3. Dorsman J. C., Doorenbosch M. M., Maurer C. T., de Winde J. H., Mager W. H., Planta R. J., Grivell L. A. An ARS/silencer binding factor also activates two ribosomal protein genes in yeast. Nucleic Acids Res. 1989 Jul 11;17(13):4917–4923. doi: 10.1093/nar/17.13.4917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eng F. J., Warner J. R. Structural basis for the regulation of splicing of a yeast messenger RNA. Cell. 1991 May 31;65(5):797–804. doi: 10.1016/0092-8674(91)90387-e. [DOI] [PubMed] [Google Scholar]
  5. Hamil K. G., Nam H. G., Fried H. M. Constitutive transcription of yeast ribosomal protein gene TCM1 is promoted by uncommon cis- and trans-acting elements. Mol Cell Biol. 1988 Oct;8(10):4328–4341. doi: 10.1128/mcb.8.10.4328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hardy C. F., Balderes D., Shore D. Dissection of a carboxy-terminal region of the yeast regulatory protein RAP1 with effects on both transcriptional activation and silencing. Mol Cell Biol. 1992 Mar;12(3):1209–1217. doi: 10.1128/mcb.12.3.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Henry Y. A., Chambers A., Tsang J. S., Kingsman A. J., Kingsman S. M. Characterisation of the DNA binding domain of the yeast RAP1 protein. Nucleic Acids Res. 1990 May 11;18(9):2617–2623. doi: 10.1093/nar/18.9.2617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Herruer M. H., Mager W. H., Doorenbosch T. M., Wessels P. L., Wassenaar T. M., Planta R. J. The extended promoter of the gene encoding ribosomal protein S33 in yeast consists of multiple protein binding elements. Nucleic Acids Res. 1989 Sep 25;17(18):7427–7439. doi: 10.1093/nar/17.18.7427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Herruer M. H., Mager W. H., Raué H. A., Vreken P., Wilms E., Planta R. J. Mild temperature shock affects transcription of yeast ribosomal protein genes as well as the stability of their mRNAs. Nucleic Acids Res. 1988 Aug 25;16(16):7917–7929. doi: 10.1093/nar/16.16.7917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Kim C. H., Warner J. R. Mild temperature shock alters the transcription of a discrete class of Saccharomyces cerevisiae genes. Mol Cell Biol. 1983 Mar;3(3):457–465. doi: 10.1128/mcb.3.3.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klein C., Struhl K. Protein kinase A mediates growth-regulated expression of yeast ribosomal protein genes by modulating RAP1 transcriptional activity. Mol Cell Biol. 1994 Mar;14(3):1920–1928. doi: 10.1128/mcb.14.3.1920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kyrion G., Boakye K. A., Lustig A. J. C-terminal truncation of RAP1 results in the deregulation of telomere size, stability, and function in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Nov;12(11):5159–5173. doi: 10.1128/mcb.12.11.5159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Leer R. J., van Raamsdonk-Duin M. M., Hagendoorn M. J., Mager W. H., Planta R. J. Structural comparison of yeast ribosomal protein genes. Nucleic Acids Res. 1984 Sep 11;12(17):6685–6700. doi: 10.1093/nar/12.17.6685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Li B., Warner J. R. Mutation of the Rab6 homologue of Saccharomyces cerevisiae, YPT6, inhibits both early Golgi function and ribosome biosynthesis. J Biol Chem. 1996 Jul 12;271(28):16813–16819. doi: 10.1074/jbc.271.28.16813. [DOI] [PubMed] [Google Scholar]
  16. Liu C., Mao X., Lustig A. J. Mutational analysis defines a C-terminal tail domain of RAP1 essential for Telomeric silencing in Saccharomyces cerevisiae. Genetics. 1994 Dec;138(4):1025–1040. doi: 10.1093/genetics/138.4.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lupashin V. V., Waters M. G. t-SNARE activation through transient interaction with a rab-like guanosine triphosphatase. Science. 1997 May 23;276(5316):1255–1258. doi: 10.1126/science.276.5316.1255. [DOI] [PubMed] [Google Scholar]
  18. Mizuta K., Warner J. R. Continued functioning of the secretory pathway is essential for ribosome synthesis. Mol Cell Biol. 1994 Apr;14(4):2493–2502. doi: 10.1128/mcb.14.4.2493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Moretti P., Freeman K., Coodly L., Shore D. Evidence that a complex of SIR proteins interacts with the silencer and telomere-binding protein RAP1. Genes Dev. 1994 Oct 1;8(19):2257–2269. doi: 10.1101/gad.8.19.2257. [DOI] [PubMed] [Google Scholar]
  20. Mori K., Sant A., Kohno K., Normington K., Gething M. J., Sambrook J. F. A 22 bp cis-acting element is necessary and sufficient for the induction of the yeast KAR2 (BiP) gene by unfolded proteins. EMBO J. 1992 Jul;11(7):2583–2593. doi: 10.1002/j.1460-2075.1992.tb05323.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Normington K., Kohno K., Kozutsumi Y., Gething M. J., Sambrook J. S. cerevisiae encodes an essential protein homologous in sequence and function to mammalian BiP. Cell. 1989 Jun 30;57(7):1223–1236. doi: 10.1016/0092-8674(89)90059-7. [DOI] [PubMed] [Google Scholar]
  22. Ogden J. E., Stanway C., Kim S., Mellor J., Kingsman A. J., Kingsman S. M. Efficient expression of the Saccharomyces cerevisiae PGK gene depends on an upstream activation sequence but does not require TATA sequences. Mol Cell Biol. 1986 Dec;6(12):4335–4343. doi: 10.1128/mcb.6.12.4335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ossig R., Dascher C., Trepte H. H., Schmitt H. D., Gallwitz D. The yeast SLY gene products, suppressors of defects in the essential GTP-binding Ypt1 protein, may act in endoplasmic reticulum-to-Golgi transport. Mol Cell Biol. 1991 Jun;11(6):2980–2993. doi: 10.1128/mcb.11.6.2980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Planta R. J., Raué H. A. Control of ribosome biogenesis in yeast. Trends Genet. 1988 Mar;4(3):64–68. doi: 10.1016/0168-9525(88)90042-x. [DOI] [PubMed] [Google Scholar]
  25. Rotenberg M. O., Woolford J. L., Jr Tripartite upstream promoter element essential for expression of Saccharomyces cerevisiae ribosomal protein genes. Mol Cell Biol. 1986 Feb;6(2):674–687. doi: 10.1128/mcb.6.2.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schwindinger W. F., Warner J. R. Transcriptional elements of the yeast ribosomal protein gene CYH2. J Biol Chem. 1987 Apr 25;262(12):5690–5695. [PubMed] [Google Scholar]
  27. Shampay J., Blackburn E. H. Generation of telomere-length heterogeneity in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1988 Jan;85(2):534–538. doi: 10.1073/pnas.85.2.534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shen W. C., Green M. R. Yeast TAF(II)145 functions as a core promoter selectivity factor, not a general coactivator. Cell. 1997 Aug 22;90(4):615–624. doi: 10.1016/s0092-8674(00)80523-1. [DOI] [PubMed] [Google Scholar]
  29. Shore D., Nasmyth K. Purification and cloning of a DNA binding protein from yeast that binds to both silencer and activator elements. Cell. 1987 Dec 4;51(5):721–732. doi: 10.1016/0092-8674(87)90095-x. [DOI] [PubMed] [Google Scholar]
  30. Shore D. RAP1: a protean regulator in yeast. Trends Genet. 1994 Nov;10(11):408–412. doi: 10.1016/0168-9525(94)90058-2. [DOI] [PubMed] [Google Scholar]
  31. Shuai K., Warner J. R. A temperature sensitive mutant of Saccharomyces cerevisiae defective in pre-rRNA processing. Nucleic Acids Res. 1991 Sep 25;19(18):5059–5064. doi: 10.1093/nar/19.18.5059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sussel L., Shore D. Separation of transcriptional activation and silencing functions of the RAP1-encoded repressor/activator protein 1: isolation of viable mutants affecting both silencing and telomere length. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7749–7753. doi: 10.1073/pnas.88.17.7749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tsang J. S., Henry Y. A., Chambers A., Kingsman A. J., Kingsman S. M. Phosphorylation influences the binding of the yeast RAP1 protein to the upstream activating sequence of the PGK gene. Nucleic Acids Res. 1990 Dec 25;18(24):7331–7337. doi: 10.1093/nar/18.24.7331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Velculescu V. E., Zhang L., Zhou W., Vogelstein J., Basrai M. A., Bassett D. E., Jr, Hieter P., Vogelstein B., Kinzler K. W. Characterization of the yeast transcriptome. Cell. 1997 Jan 24;88(2):243–251. doi: 10.1016/s0092-8674(00)81845-0. [DOI] [PubMed] [Google Scholar]
  35. Verdier J. M., Stalder R., Roberge M., Amati B., Sentenac A., Gasser S. M. Preparation and characterization of yeast nuclear extracts for efficient RNA polymerase B (II)-dependent transcription in vitro. Nucleic Acids Res. 1990 Dec 11;18(23):7033–7039. doi: 10.1093/nar/18.23.7033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Woudt L. P., Smit A. B., Mager W. H., Planta R. J. Conserved sequence elements upstream of the gene encoding yeast ribosomal protein L25 are involved in transcription activation. EMBO J. 1986 May;5(5):1037–1040. doi: 10.1002/j.1460-2075.1986.tb04319.x. [DOI] [PMC free article] [PubMed] [Google Scholar]