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A putative stimulatory role for activator turnover in gene expression

Nature volume 438, pages 113–116 (2005)Cite this article

Abstract

The ubiquitin–proteasome system (UPS) promotes the destruction of target proteins by attaching to them a ubiquitin chain that is recognized by the 26S proteasome1. The UPS influences most cellular processes, and its targets include transcriptional activators that are primary determinants of gene expression. Emerging evidence indicates that non-proteolytic functions of the UPS might stimulate transcriptional activity2,3. Here we show that the proteolysis of some transcriptional activators by the UPS can stimulate their function. We focused on the role of UPS-dependent proteolysis in the function of inducible transcriptional activators in yeast, and found that inhibition of the proteasome4 reduced transcription of the targets of the activators Gcn4, Gal4 and Ino2/4. In addition, mutations in SCFCdc4, the ubiquitin ligase for Gcn4 (ref. 5), or mutations in ubiquitin that prevent degradation6, also impaired the transcription of Gcn4 targets. These transcriptional defects were manifested despite the enhanced abundance of Gcn4 on cognate promoters. Proteasome inhibition also decreased the association of RNA polymerase II with Gcn4, Gal4 and Ino2/4 targets, as did mutations in SCFCdc4 for Gcn4 targets. Expression of a stable phospho-site mutant of Gcn4 (ref. 7) or disruption of the kinases that target Gcn4 for turnover5,7 alleviated the sensitivity of Gcn4 activity to defects in the UPS.

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Figure 1: UPS-dependent proteolysis positively regulates inducible transcriptional activators.
Figure 2: Defects in the UPS lead to the accumulation of ubiquitinated Gcn4 and impair the association of RNA polymerase II with Gcn4 and Gal4 targets.
Figure 3: The UPS has little effect on the activity of stable, non-phosphorylated versions of Gcn4.

References

  1. Pickart, C. M. Back to the future with ubiquitin. Cell 116, 181–190 (2004)

    Article  CAS  Google Scholar 

  2. Muratani, M. & Tansey, W. P. How the ubiquitin–proteasome system controls transcription. Nature Rev. Mol. Cell Biol. 4, 192–201 (2003)

    Article  CAS  Google Scholar 

  3. Lipford, J. R. & Deshaies, R. J. Diverse roles for ubiquitin-dependent proteolysis in transcriptional activation. Nature Cell Biol. 5, 845–850 (2003)

    Article  CAS  Google Scholar 

  4. Lee, D. H. & Goldberg, A. L. Proteasome inhibitors: valuable new tools for cell biologists. Trends Cell Biol. 8, 397–403 (1998)

    Article  CAS  Google Scholar 

  5. Meimoun, A. et al. Degradation of the transcription factor Gcn4 requires the kinase Pho85 and the SCF(CDC4) ubiquitin-ligase complex. Mol. Biol. Cell 11, 915–927 (2000)

    Article  CAS  Google Scholar 

  6. Finley, D. et al. Inhibition of proteolysis and cell-cycle progression in a multiubiquitination-deficient yeast mutant. Mol. Cell. Biol. 14, 5501–5509 (1994)

    Article  CAS  Google Scholar 

  7. Chi, Y. et al. Negative regulation of Gcn4 and Msn2 transcription factors by Srb10 cyclin-dependent kinase. Genes Dev. 15, 1078–1092 (2001)

    Article  CAS  Google Scholar 

  8. Adams, J. & Kauffman, M. Development of the proteasome inhibitor Velcade (bortezomib). Cancer Invest. 22, 304–311 (2004)

    Article  CAS  Google Scholar 

  9. Mitsiades, N. et al. Molecular sequelae of proteasome inhibition in human multiple myeloma cells. Proc. Natl Acad. Sci. USA 99, 14374–14379 (2002)

    Article  ADS  CAS  Google Scholar 

  10. Fleming, J. A. et al. Complementary whole-genome technologies reveal the cellular response to proteasome inhibition by PS-341. Proc. Natl Acad. Sci. USA 99, 1461–1466 (2002)

    Article  ADS  CAS  Google Scholar 

  11. Hinnebusch, A. G. & Natarajan, K. Gcn4p, a master regulator of gene expression, is controlled at multiple levels by diverse signals of starvation and stress. Eukaryot. Cell 1, 22–32 (2002)

    Article  CAS  Google Scholar 

  12. Kornitzer, D., Raboy, B., Kulka, R. G. & Fink, G. R. Regulated degradation of the transcription factor GCN4. EMBO J. 13, 6021–6030 (1994)

    Article  CAS  Google Scholar 

  13. Albrecht, G., Mosch, H. U., Hoffmann, B., Reusser, U. & Braus, G. H. Monitoring the Gcn4 protein-mediated response in the yeast Saccharomyces cerevisiae. J. Biol. Chem. 273, 12696–12702 (1998)

    Article  CAS  Google Scholar 

  14. Hilt, W., Enenkel, C., Gruhler, A., Singer, T. & Wolf, D. H. The PRE4 gene codes for a subunit of the yeast proteasome necessary for peptidylglutamyl-peptide-hydrolyzing activity. Mutations link the proteasome to stress- and ubiquitin-dependent proteolysis. J. Biol. Chem. 268, 3479–3486 (1993)

    CAS  PubMed  Google Scholar 

  15. Salghetti, S. E., Caudy, A. A., Chenoweth, J. G. & Tansey, W. P. Regulation of transcriptional activation domain function by ubiquitin. Science 293, 1651–1653 (2001)

    Article  ADS  CAS  Google Scholar 

  16. Kim, S., Herbst, A., Tworkowski, K., Salghetti, S. & Tansey, W. Skp2 regulates Myc protein stability and activity. Mol. Cell 11, 1177–1188 (2003)

    Article  CAS  Google Scholar 

  17. von der Lehr, N. et al. The F-Box protein Skp2 participates in c-Myc proteosomal degradation and acts as a cofactor for c-Myc-regulated transcription. Mol. Cell 11, 1177–1188 (2003)

    Article  Google Scholar 

  18. Aparicio, O., Geisberg, J. & Struhl, K. in Current Protocols in Molecular Biology (eds Ausubel, F. M. et al.) 21.3.1–21.3.12 (Wiley, New York, 2004)

    Google Scholar 

  19. Stitzel, M. L., Durso, R. & Reese, J. C. The proteasome regulates the UV-induced activation of the AP-1-like transcription factor Gcn4. Genes Dev. 15, 128–133 (2001)

    Article  CAS  Google Scholar 

  20. Reid, G. et al. Cyclic, proteasome-mediated turnover of unliganded and liganded ERα on responsive promoters is an integral feature of estrogen signalling. Mol. Cell 11, 695–707 (2003)

    Article  CAS  Google Scholar 

  21. Gonzalez, F., Delahodde, A., Kodadek, T. & Johnston, S. A. Recruitment of a 19S proteasome subcomplex to an activated promoter. Science 296, 548–550 (2002)

    Article  ADS  CAS  Google Scholar 

  22. Morris, M. C. et al. Cks1-dependent proteasome recruitment and activation of CDC20 transcription in budding yeast. Nature 423, 1009–1013 (2003)

    Article  ADS  CAS  Google Scholar 

  23. Verma, R. et al. Deubiquitination and degradation of proteins by the 26S proteasome requires the Rpn11 metalloprotease motif. Science 298, 611–615 (2002)

    Article  ADS  CAS  Google Scholar 

  24. Muratani, M., Kung, C., Shokat, K. M. & Tansey, W. P. The F box protein Dsg1/Mdm30 is a transcriptional coactivator that stimulates Gal4 turnover and cotranscriptional mRNA processing. Cell 120, 887–899 (2005)

    Article  CAS  Google Scholar 

  25. Hirst, M., Kobor, M. S., Kuriakose, N., Greenblatt, J. & Sadowski, I. GAL4 is regulated by the RNA polymerase II holoenzyme-associated cyclin-dependent protein kinase SRB10/CDK8. Mol. Cell 3, 673–678 (1999)

    Article  CAS  Google Scholar 

  26. Guthrie, C. & Fink, G. R. Guide to Yeast Genetics and Molecular Biology (Academic, San Diego, 1991)

    Google Scholar 

  27. Drysdale, C. M. et al. The transcriptional activator GCN4 contains multiple activation domains that are critically dependent on hydrophobic amino acids. Mol. Cell. Biol. 15, 1220–1233 (1995)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank D. Finley, D. H. Wolf, R. Young, A. Hinnebusch, J. Shaw, B. Westermann, J. Nunnari and G. Braus for gifts of strains and reagents; B. Tansey for communicating results before publication; and J. Shaw, B. Westermann, J. Nunnari, S. Sadis, A. Ansari and the members of the Deshaies laboratory for comments and criticism. This work was supported in part by an NIH Research Project Grant to R.J.D. R.J.D. is an Investigator of the Howard Hughes Medical Institute. J.R.L. was supported by an NIH National Research Service Award and a Caltech-Amgen Postdoctoral Fellowship.

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  1. Yong Chi

    Present address: Institute for Systems Biology, 1441 North 34th Street, Seattle, Washington, 98103, USA

Authors and Affiliations

  1. Howard Hughes Medical Institute, Division of Biology, MC 156-29, California Institute of Technology, 1200 E. California Boulevard, California, 91125, Pasadena, USA

    J. Russell Lipford, Geoffrey T. Smith, Yong Chi & Raymond J. Deshaies

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  1. J. Russell Lipford
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  2. Geoffrey T. Smith
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Correspondence to Raymond J. Deshaies.

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Supplementary information

Supplementary Notes

This file contains Supplementary Table 1 and 2, Supplementary Figure Legends, Supplementary Methods, and Supplementary Discussion. This file also contains details of NIH support. (DOC 166 kb)

Supplementary Figure 1

Proteasome inhibition represses the transcription of many Gcn4 targets and of INO1, a target of the transcriptional activators, Ino2 and Ino4. (PDF 36 kb)

Supplementary Figure 2

Gal4-TAP functions similarly to wild-type Gal4. pdr5δ, GAL4-TAP strains were treated as in Fig. 1b. (PDF 29 kb)

Supplementary Figure 3

Proteasome inhibition does not affect the dynamic association of Gal80 with a Gal4 target. (PDF 29 kb)

Supplementary Figure 4

Gcn4 is required for the association of RNA polymerase II (polII) with HIS4. (PDF 16 kb)

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Lipford, J., Smith, G., Chi, Y. et al. A putative stimulatory role for activator turnover in gene expression. Nature 438, 113–116 (2005). https://doi.org/10.1038/nature04098

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