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PMID: 9851990 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Suppression of an Hsp70 mutant phenotype in Saccharomyces cerevisiae through loss of function of the chromatin component Sin1p/Spt2p.

Journal of bacteriology ·Vol. 180 ·No. 24 ·1998-12-00 ·Pages 6484-92

Baxter BK, Craig EA

Abstract

The Ssa subfamily of Hsp70 molecular chaperones in the budding yeast Saccharomyces cerevisiae has four members, encoded by SSA1, SSA2, SSA3, and SSA4. Deletion of the two constitutively expressed genes, SSA1 and SSA2, results in cells which are slow growing and temperature sensitive. In this study, we demonstrate that an extragenic suppressor of the temperature sensitivity of ssa1 ssa2 strains, EXA1-1, is a loss-of-function mutation in SIN1/SPT2, which encodes a nonhistone component of chromatin. Loss of function of Sin1p leads to overexpression of SSA3 in the ssa1 ssa2 mutant background, at a level which is sufficient to mediate suppression. In a strain which is wild type for SSA genes, we detected no effect of Sin1p on Ssa3p expression except under conditions of heat shock. Existing data indicate that expression of SSA3 in the ssa1 ssa2 mutant background as well as in heat-shocked wild-type strains is mediated by the heat shock transcription factor HSF. Our findings suggest that it is HSF-mediated induction of SSA3 which is modulated by Sin1p. The EXA1-1 suppressor mutation thus improves the growth of ssa1 ssa2 strains by selectively increasing HSF-mediated expression of SSA3.

MeSH Terms
Bacterial Proteins/biosynthesis Chromatin Chromosomal Proteins, Non-Histone/genetics,physiology DNA-Binding Proteins/genetics,physiology Gene Expression HSP40 Heat-Shock Proteins HSP70 Heat-Shock Proteins/biosynthesis,genetics Heat-Shock Proteins/biosynthesis Mutagenesis Phenotype Saccharomyces cerevisiae/genetics,physiology Saccharomyces cerevisiae Proteins Suppression, Genetic
Chemicals
Bacterial Proteins Chromatin Chromosomal Proteins, Non-Histone DNA-Binding Proteins HSP40 Heat-Shock Proteins HSP70 Heat-Shock Proteins Heat-Shock Proteins SSA3 protein, S cerevisiae Saccharomyces cerevisiae Proteins Streptomyces trypsin inhibitor 1 protein, Streptomyces YDJ1 protein, S cerevisiae HsP104 protein, S cerevisiae SPT2 protein, S cerevisiae
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Baxter B K
Department of Biomolecular Chemistry, University of Wisconsin, Madison, Wisconsin 53706, USA.
Craig E A
References (38)
38 references, click to expand
  1. Pedigree analysis of plasmid segregation in yeast.
    Cell. 1983 Oct;34(3):961-70 PMID: 6354471
  2. Protein folding in vivo: unraveling complex pathways.
    Cell. 1997 Jul 25;90(2):201-4 PMID: 9244293
  3. Mutations of the heat inducible 70 kilodalton genes of yeast confer temperature sensitive growth.
    Cell. 1984 Oct;38(3):841-9 PMID: 6386178
  4. Isolation and characterization of the SPT2 gene, a negative regulator of Ty-controlled yeast gene expression.
    Mol Cell Biol. 1985 Jul;5(7):1543-53 PMID: 2991744
  5. Activation of the yeast HO gene by release from multiple negative controls.
    Cell. 1987 Feb 27;48(4):567-77 PMID: 3545494
  6. Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 May;7(5):1906-16 PMID: 3037338
  7. Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 Jul;7(7):2568-77 PMID: 3302682
  8. A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides.
    Nature. 1988 Apr 28;332(6167):800-5 PMID: 3282178
  9. 70K heat shock related proteins stimulate protein translocation into microsomes.
    Nature. 1988 Apr 28;332(6167):805-10 PMID: 3282179
  10. Yeast/E. coli shuttle vectors with multiple unique restriction sites.
    Yeast. 1986 Sep;2(3):163-7 PMID: 3333305
  11. Yeast Hsp70 RNA levels vary in response to the physiological status of the cell.
    J Bacteriol. 1989 May;171(5):2680-8 PMID: 2651414
  12. Initiation of lambda DNA replication with purified host- and bacteriophage-encoded proteins: the role of the dnaK, dnaJ and grpE heat shock proteins.
    EMBO J. 1989 May;8(5):1601-8 PMID: 2527744
  13. Isolation and characterization of STI1, a stress-inducible gene from Saccharomyces cerevisiae.
    Mol Cell Biol. 1989 Sep;9(9):3638-46 PMID: 2674681
  14. Self-regulation of 70-kilodalton heat shock proteins in Saccharomyces cerevisiae.
    Mol Cell Biol. 1990 Apr;10(4):1622-32 PMID: 2181281
  15. Transcriptional regulation of SSA3, an HSP70 gene from Saccharomyces cerevisiae.
    Mol Cell Biol. 1990 Jun;10(6):3262-7 PMID: 2188113
  16. A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae.
    Nucleic Acids Res. 1990 May 25;18(10):3091-2 PMID: 2190191
  17. Regulation of a yeast HSP70 gene by a cAMP responsive transcriptional control element.
    EMBO J. 1990 Aug;9(8):2543-53 PMID: 1695149
  18. Structure and regulation of the SSA4 HSP70 gene of Saccharomyces cerevisiae.
    J Biol Chem. 1990 Nov 5;265(31):18912-21 PMID: 2121731
  19. A functional interaction between the C-terminal domain of RNA polymerase II and the negative regulator SIN1.
    Cell. 1991 Mar 22;64(6):1135-43 PMID: 2004420
  20. A negative regulator of HO transcription, SIN1 (SPT2), is a nonspecific DNA-binding protein related to HMG1.
    Mol Cell Biol. 1991 Aug;11(8):4135-46 PMID: 2072912
  21. Characterization of YDJ1: a yeast homologue of the bacterial dnaJ protein.
    J Cell Biol. 1991 Aug;114(4):609-21 PMID: 1869583
  22. MAS5, a yeast homolog of DnaJ involved in mitochondrial protein import.
    Mol Cell Biol. 1992 Jan;12(1):283-91 PMID: 1729605
  23. Isolation and characterization of extragenic suppressors of mutations in the SSA hsp70 genes of Saccharomyces cerevisiae.
    Genetics. 1992 Jun;131(2):277-85 PMID: 1644272
  24. New vectors for high level expression of recombinant proteins in bacteria.
    Anal Biochem. 1992 May 1;202(2):293-8 PMID: 1519755
  25. Evidence that SNF2/SWI2 and SNF5 activate transcription in yeast by altering chromatin structure.
    Genes Dev. 1992 Dec;6(12A):2288-98 PMID: 1459453
  26. YDJ1p facilitates polypeptide translocation across different intracellular membranes by a conserved mechanism.
    Cell. 1992 Dec 24;71(7):1143-55 PMID: 1473150
  27. Mutational and functional analysis of dominant SPT2 (SIN1) suppressor alleles in Saccharomyces cerevisiae.
    Mol Cell Biol. 1993 Sep;13(9):5393-407 PMID: 8395004
  28. Genetic evidence for a functional relationship between Hsp104 and Hsp70.
    J Bacteriol. 1993 Oct;175(20):6484-91 PMID: 8407824
  29. Nucleosome disruption and enhancement of activator binding by a human SW1/SNF complex.
    Nature. 1994 Aug 11;370(6489):477-81 PMID: 8047169
  30. Facilitated binding of TATA-binding protein to nucleosomal DNA.
    Nature. 1994 Aug 11;370(6489):481-5 PMID: 8047170
  31. Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds.
    Gene. 1995 Apr 14;156(1):119-22 PMID: 7737504
  32. Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure.
    Yeast. 1995 Apr 15;11(4):355-60 PMID: 7785336
  33. Conserved ATPase and luciferase refolding activities between bacteria and yeast Hsp70 chaperones and modulators.
    FEBS Lett. 1995 Jul 24;368(3):435-40 PMID: 7635193
  34. A heat shock transcription factor with reduced activity suppresses a yeast HSP70 mutant.
    Mol Cell Biol. 1995 Sep;15(9):4890-7 PMID: 7651408
  35. Persistent site-specific remodeling of a nucleosome array by transient action of the SWI/SNF complex.
    Science. 1996 Jul 26;273(5274):513-6 PMID: 8662543
  36. Functional interaction of cytosolic hsp70 and a DnaJ-related protein, Ydj1p, in protein translocation in vivo.
    Mol Cell Biol. 1996 Aug;16(8):4378-86 PMID: 8754838
  37. The yeast SWI-SNF complex facilitates binding of a transcriptional activator to nucleosomal sites in vivo.
    Mol Cell Biol. 1997 Aug;17(8):4811-9 PMID: 9234737
  38. Mutations affecting Ty-mediated expression of the HIS4 gene of Saccharomyces cerevisiae.
    Genetics. 1984 Jun;107(2):179-97 PMID: 6329902
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
Published
1998-12-00
Pages
6484-92
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC107749
Subset
IM
Grants
NIGMS NIH HHS · R01 GM031107 · United States
NIGMS NIH HHS · R37 GM031107 · United States
NIGMS NIH HHS · GM31107 · United States
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