Skip to main content
PMC home page
Genetics logoLink to Genetics
. 2000 Oct;156(2):559–570. doi: 10.1093/genetics/156.2.559

A role for cytosolic hsp70 in yeast [PSI(+)] prion propagation and [PSI(+)] as a cellular stress.

G Jung 1, G Jones 1, R D Wegrzyn 1, D C Masison 1
PMCID: PMC1461277  PMID: 11014806

Abstract

[PSI(+)] is a prion (infectious protein) of Sup35p, a subunit of the Saccharomyces cerevisiae translation termination factor. We isolated a dominant allele, SSA1-21, of a gene encoding an Hsp70 chaperone that impairs [PSI(+)] mitotic stability and weakens allosuppression caused by [PSI(+)]. While [PSI(+)] stability is normal in strains lacking SSA1, SSA2, or both, SSA1-21 strains with a deletion of SSA2 cannot propagate [PSI(+)]. SSA1-21 [PSI(+)] strains are hypersensitive to curing of [PSI(+)] by guanidine-hydrochloride and partially cured of [PSI(+)] by rapid induction of the heat-shock response but not by growth at 37 degrees. The number of inheritable [PSI(+)] particles is significantly reduced in SSA1-21 cells. SSA1-21 effects on [PSI(+)] appear to be independent of Hsp104, another stress-inducible protein chaperone known to be involved in [PSI(+)] propagation. We propose that cytosolic Hsp70 is important for the formation of Sup35p polymers characteristic of [PSI(+)] from preexisting material and that Ssa1-21p both lacks and interferes with this activity. We further demonstrate that the negative effect of heat stress on [PSI(+)] phenotype directly correlates with solubility of Sup35p and find that in wild-type strains the presence of [PSI(+)] causes a stress that elevates basal expression of Hsp104 and SSA1.

Full Text

The Full Text of this article is available as a PDF (407.5 KB).

Selected References

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

  1. Bonetti B., Fu L., Moon J., Bedwell D. M. The efficiency of translation termination is determined by a synergistic interplay between upstream and downstream sequences in Saccharomyces cerevisiae. J Mol Biol. 1995 Aug 18;251(3):334–345. doi: 10.1006/jmbi.1995.0438. [DOI] [PubMed] [Google Scholar]
  2. Boorstein W. R., Craig E. A. Structure and regulation of the SSA4 HSP70 gene of Saccharomyces cerevisiae. J Biol Chem. 1990 Nov 5;265(31):18912–18921. [PubMed] [Google Scholar]
  3. Boorstein W. R., Ziegelhoffer T., Craig E. A. Molecular evolution of the HSP70 multigene family. J Mol Evol. 1994 Jan;38(1):1–17. doi: 10.1007/BF00175490. [DOI] [PubMed] [Google Scholar]
  4. Chernoff Y. O., Lindquist S. L., Ono B., Inge-Vechtomov S. G., Liebman S. W. Role of the chaperone protein Hsp104 in propagation of the yeast prion-like factor [psi+]. Science. 1995 May 12;268(5212):880–884. doi: 10.1126/science.7754373. [DOI] [PubMed] [Google Scholar]
  5. Conde J., Fink G. R. A mutant of Saccharomyces cerevisiae defective for nuclear fusion. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3651–3655. doi: 10.1073/pnas.73.10.3651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cox B. S., Tuite M. F., McLaughlin C. S. The psi factor of yeast: a problem in inheritance. Yeast. 1988 Sep;4(3):159–178. doi: 10.1002/yea.320040302. [DOI] [PubMed] [Google Scholar]
  7. Craig E. A., Gross C. A. Is hsp70 the cellular thermometer? Trends Biochem Sci. 1991 Apr;16(4):135–140. doi: 10.1016/0968-0004(91)90055-z. [DOI] [PubMed] [Google Scholar]
  8. DePace A. H., Santoso A., Hillner P., Weissman J. S. A critical role for amino-terminal glutamine/asparagine repeats in the formation and propagation of a yeast prion. Cell. 1998 Jun 26;93(7):1241–1252. doi: 10.1016/s0092-8674(00)81467-1. [DOI] [PubMed] [Google Scholar]
  9. Derkatch I. L., Chernoff Y. O., Kushnirov V. V., Inge-Vechtomov S. G., Liebman S. W. Genesis and variability of [PSI] prion factors in Saccharomyces cerevisiae. Genetics. 1996 Dec;144(4):1375–1386. doi: 10.1093/genetics/144.4.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eaglestone S. S., Cox B. S., Tuite M. F. Translation termination efficiency can be regulated in Saccharomyces cerevisiae by environmental stress through a prion-mediated mechanism. EMBO J. 1999 Apr 1;18(7):1974–1981. doi: 10.1093/emboj/18.7.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eaglestone S. S., Ruddock L. W., Cox B. S., Tuite M. F. Guanidine hydrochloride blocks a critical step in the propagation of the prion-like determinant [PSI(+)] of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 2000 Jan 4;97(1):240–244. doi: 10.1073/pnas.97.1.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glover J. R., Kowal A. S., Schirmer E. C., Patino M. M., Liu J. J., Lindquist S. Self-seeded fibers formed by Sup35, the protein determinant of [PSI+], a heritable prion-like factor of S. cerevisiae. Cell. 1997 May 30;89(5):811–819. doi: 10.1016/s0092-8674(00)80264-0. [DOI] [PubMed] [Google Scholar]
  13. Grant C. M., Firoozan M., Tuite M. F. Mistranslation induces the heat-shock response in the yeast Saccharomyces cerevisiae. Mol Microbiol. 1989 Feb;3(2):215–220. doi: 10.1111/j.1365-2958.1989.tb01810.x. [DOI] [PubMed] [Google Scholar]
  14. Guarente L. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 1983;101:181–191. doi: 10.1016/0076-6879(83)01013-7. [DOI] [PubMed] [Google Scholar]
  15. Ingolia T. D., Slater M. R., Craig E. A. Saccharomyces cerevisiae contains a complex multigene family related to the major heat shock-inducible gene of Drosophila. Mol Cell Biol. 1982 Nov;2(11):1388–1398. doi: 10.1128/mcb.2.11.1388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. King C. Y., Tittmann P., Gross H., Gebert R., Aebi M., Wüthrich K. Prion-inducing domain 2-114 of yeast Sup35 protein transforms in vitro into amyloid-like filaments. Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6618–6622. doi: 10.1073/pnas.94.13.6618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lawrence C. W. Classical mutagenesis techniques. Methods Enzymol. 1991;194:273–281. doi: 10.1016/0076-6879(91)94021-4. [DOI] [PubMed] [Google Scholar]
  18. Liebman S. W., Stewart J. W., Sherman F. Serine substitutions caused by an ochre suppressor in yeast. J Mol Biol. 1975 Jun 5;94(4):595–610. doi: 10.1016/0022-2836(75)90324-1. [DOI] [PubMed] [Google Scholar]
  19. Lin B, Shih MC, Bohanon TM, Ice GE, Dutta P. Phase diagram of a lipid monolayer on the surface of water. Phys Rev Lett. 1990 Jul 9;65(2):191–194. doi: 10.1103/PhysRevLett.65.191. [DOI] [PubMed] [Google Scholar]
  20. Masison D. C., Maddelein M. L., Wickner R. B. The prion model for [URE3] of yeast: spontaneous generation and requirements for propagation. Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12503–12508. doi: 10.1073/pnas.94.23.12503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McCready S. J., Cox B. S., McLaughlin C. S. The extrachromosomal control of nonsense suppression in yeast: an analysis of the elimination of [psi+] in the presence of a nuclear gene PNM. Mol Gen Genet. 1977 Feb 15;150(3):265–270. doi: 10.1007/BF00268125. [DOI] [PubMed] [Google Scholar]
  22. Newnam G. P., Wegrzyn R. D., Lindquist S. L., Chernoff Y. O. Antagonistic interactions between yeast chaperones Hsp104 and Hsp70 in prion curing. Mol Cell Biol. 1999 Feb;19(2):1325–1333. doi: 10.1128/mcb.19.2.1325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nicolet C. M., Craig E. A. Functional analysis of a conserved amino-terminal region of HSP70 by site-directed mutagenesis. Yeast. 1991 Oct;7(7):699–716. doi: 10.1002/yea.320070706. [DOI] [PubMed] [Google Scholar]
  24. Oka M., Nakai M., Endo T., Lim C. R., Kimata Y., Kohno K. Loss of Hsp70-Hsp40 chaperone activity causes abnormal nuclear distribution and aberrant microtubule formation in M-phase of Saccharomyces cerevisiae. J Biol Chem. 1998 Nov 6;273(45):29727–29737. doi: 10.1074/jbc.273.45.29727. [DOI] [PubMed] [Google Scholar]
  25. Parsell D. A., Kowal A. S., Singer M. A., Lindquist S. Protein disaggregation mediated by heat-shock protein Hsp104. Nature. 1994 Dec 1;372(6505):475–478. doi: 10.1038/372475a0. [DOI] [PubMed] [Google Scholar]
  26. Patino M. M., Liu J. J., Glover J. R., Lindquist S. Support for the prion hypothesis for inheritance of a phenotypic trait in yeast. Science. 1996 Aug 2;273(5275):622–626. doi: 10.1126/science.273.5275.622. [DOI] [PubMed] [Google Scholar]
  27. Paushkin S. V., Kushnirov V. V., Smirnov V. N., Ter-Avanesyan M. D. In vitro propagation of the prion-like state of yeast Sup35 protein. Science. 1997 Jul 18;277(5324):381–383. doi: 10.1126/science.277.5324.381. [DOI] [PubMed] [Google Scholar]
  28. Paushkin S. V., Kushnirov V. V., Smirnov V. N., Ter-Avanesyan M. D. Propagation of the yeast prion-like [psi+] determinant is mediated by oligomerization of the SUP35-encoded polypeptide chain release factor. EMBO J. 1996 Jun 17;15(12):3127–3134. [PMC free article] [PubMed] [Google Scholar]
  29. Rose M. D., Novick P., Thomas J. H., Botstein D., Fink G. R. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. doi: 10.1016/0378-1119(87)90232-0. [DOI] [PubMed] [Google Scholar]
  30. Sherman F. Getting started with yeast. Methods Enzymol. 1991;194:3–21. doi: 10.1016/0076-6879(91)94004-v. [DOI] [PubMed] [Google Scholar]
  31. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tatzelt J., Zuo J., Voellmy R., Scott M., Hartl U., Prusiner S. B., Welch W. J. Scrapie prions selectively modify the stress response in neuroblastoma cells. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2944–2948. doi: 10.1073/pnas.92.7.2944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tuite M. F., Mundy C. R., Cox B. S. Agents that cause a high frequency of genetic change from [psi+] to [psi-] in Saccharomyces cerevisiae. Genetics. 1981 Aug;98(4):691–711. doi: 10.1093/genetics/98.4.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Werner-Washburne M., Becker J., Kosic-Smithers J., Craig E. A. Yeast Hsp70 RNA levels vary in response to the physiological status of the cell. J Bacteriol. 1989 May;171(5):2680–2688. doi: 10.1128/jb.171.5.2680-2688.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Werner-Washburne M., Craig E. A. Expression of members of the Saccharomyces cerevisiae hsp70 multigene family. Genome. 1989;31(2):684–689. doi: 10.1139/g89-125. [DOI] [PubMed] [Google Scholar]
  36. Werner-Washburne M., Stone D. E., Craig E. A. Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Jul;7(7):2568–2577. doi: 10.1128/mcb.7.7.2568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wickner R. B. [URE3] as an altered URE2 protein: evidence for a prion analog in Saccharomyces cerevisiae. Science. 1994 Apr 22;264(5158):566–569. doi: 10.1126/science.7909170. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES