Complex interactions among members of an essential subfamily of hsp70 genes in Saccharomyces cerevisiae (original) (raw)

1987, Molecular and cellular biology

Abstract

Saccharomyces cerevisiae contains a large family of genes related to hsp70, the major heat shock-inducible gene of Drosophila melanogaster. One subfamily, identified by sequence homology, contains four genes, SSA1, SSA2, SSA3, and SSA4 (formerly YG100, YG102, YG106, and YG107, respectively). Previous studies showed that strains containing mutations in SSA1 and SSA2 are temperature sensitive for growth. SSA4, which is normally heat inducible and not expressed during vegetative growth, is expressed at high levels in ssa1 ssa2 strains at 23 degrees C. We constructed mutations in SSA3 and SSA4 and analyzed strains carrying mutations in the four genes. Strains carrying mutations in SSA3 SSA4 or SSA3 and SSA4 were indistinguishable from the wild type. However, ssa1 ssa2 ssa4 strains were inviable. SSA3, like SSA4, is a heat-inducible gene that is not normally expressed at 23 degrees C. Nevertheless, an intact copy of SSA3 regulated by the constitutive SSA2 promoter was capable of rescuing...

Loading...

Loading Preview

Sorry, preview is currently unavailable. You can download the paper by clicking the button above.

References (37)

  1. MATa-leu2b ade2 Atrpl ura3 Iys2 T153a.... MATot-leu2 ade2 his3 lys2
  2. DS13a .... MATot-leu2 Atrpl ura3 his3 Iys2 GAL2 MW11C .... MATa-leu2 Atrpl ura3 ade2 lys2 (ssa4::URA3) MW14 .... MATa-leu2 Atrpl ura3 lys2 (ssa3::TRPI)
  3. MW42 .... MATa-Ieu2 Atrpl ura3 Iys2 his3 (ssa3::TRPI) (ssa4::URA3)
  4. MW63d .... MATa-leu2 Atrpl ura3 his3 Iys2 GAL2 (ssal::HIS3) (ssa2::LEU2) MATa-leu2 Atrpl ura3 lys2 his3 (ssa3::TRPI) (ssa4::URA3)
  5. MW79d .... MATa-leu2 Atrpl ura3 his3 Iys2 GAL2 (ssal::HIS3) (ssa2::LEU2)
  6. MATa-leu2 Atrpl ura3 Iys2 his3 (ssa3::TRPI) (ssa4::URA3)
  7. MW116C .... MATa-Ieu2 Atrpl ura3 his3 Iys2 (ssal::HIS3) (ssa2::LEU2) (ssa3::TRPI)
  8. MW121e.... MATa-1eu2 Atrpl ura3 his3 Iys2 (ssal::HIS3) (ssa3::TRPI) [saa4::URA3] MW122C .... MAToa-leu2 Atrpl ura3 his3 lys2 (ssa4::URA3)
  9. MW127C .... MATa-Ieu2 Atrpl ura3 his3 lys2 (ssa3::TRPI)
  10. MW13lf .... MATa-Ieu2 Atrpi ura3 his3 Iys2 GAL2 (ssal::HIS3) (ssa2::LEU2) MATa-leu2 Atrpl ura3 his3 lys2 (ssal::HIS3) (ssa4::URA3) GALI-SSAI::TRPI MW133f .... MATa-leu2 Atrpi ura3 his3 Iys2 GAL2 (ssal::HIS3) (ssa2::LEU2) MATa-leu2 Atrpi ura3 his3 Iys2 (ssal::HIS3) (ssa4::URA3) GALJ-SSAJ::TRPI MW142 .... MATa-leu2 Atrpl ura3 his3 lys2 GAL2 (ssald::HIS3) (ssa2d::URA3)
  11. MATa-leu2 Atrpi ura3 his3 lys2 GAL2 (ssal::HIS3) (ssa2::LEU2) pYe(CEN3)30::SSA2pSSA3
  12. MATa-Ieu2 Atrpl ura3 his3 lys2 (ssal::HIS3) (ssa4::URA3)
  13. MATa-leu2 Atrpl ura3 his3 Iys2 (ssal::HIS3) (ssa2::LEU2) (ssa4::URA3) pYe(CEN3)30::SSA2pSSA3 a Derived from strains obtained from M. Culbertson.
  14. Chappell, T. G., W. J. Welch, D. M. Schlossman, K. B. Palter, M. J. Schlesinger, and J. E. Rothman. 1986. Uncoating ATPase is a member of the 70 kilodalton family of stress proteins. Cell 45:3-13.
  15. Cohen, S. N., A. C. Y. Chang, and L. Hsu. 1972. Nonchromo- somal antibiotic resistance in bacteria: general transformation of Escherichia coli by R-factor DNA. Proc. Natl. Acad. Sci. USA 69:2110-2114.
  16. Craig, E. A. 1985. The heat shock response. Crit. Rev. Bio- chem. 18:239-280.
  17. Craig, E. A., and K. Jacobsen. 1984. Mutations of the heat- inducible 70 kilodalton genes of yeast confer temperature sen- sitive growth. Cell 38:841-849.
  18. Craig, E. A., and K. Jacobsen. 1985. Mutations in cognate genes of Saccharomyces cerevisiae hsp7O result in reduced growth rates at low temperatures. Mol. Cell. Biol. 5:3517-3524.
  19. Craig, E. A., M. R. Slater, W. R. Boorstein, and K. Palter. 1985. Expression of the S. cerevisiae Hsp7O multigene family, p. 659-667. In R. Calendar and L. Gold (ed.), Sequence specificity in transcription and translation. Alan R. Liss, Inc., New York.
  20. Davison, J., M. Heusterspreute, M. Merchez, and F. Brunel. 1984. Vectors with restriction-site banks I. pJRD158, a 3903-bp plasmid containing 28 unique cloning sites. Gene 28:311-318.
  21. Denhardt, D. 1966. A membrane filter technique for the detec- tion of complementary DNA. Biochem. Biophys. Res. Com- mun. 23:641-646.
  22. Ellwood, M. S., and E. A. Craig. 1984. Differential regulation of the 70K heat shock and related genes in Saccharomyces cere- visiae. Mol. Cell. Biol. 4:1454-1459.
  23. Ingolia, T. D., M. R. Slater, and E. A. Craig. 1982. Sac- charomyces cerevisiae contains a complex multigene family related to the major heat shock-inducible gene of Drosophila. Mol. Cell. Biol. 2:1388-1398.
  24. Lindquist, S. 1986. The heat-shock response. Annu. Rev. Bio- chem. 55:1151-1191.
  25. Martinez-Arias, A. E., and M. J. Casadaban. 1983. Fusion of the Saccharomyces cerevisiae leu2 gene to an Escherichia coli P-galactosidase gene. Mol. Cell. Biol. 3:580-586.
  26. Miller, M. J., N.-H. Xuong, and E. P. Geiduschek. 1979. A response of protein synthesis to temperature shift in the yeast Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 76: 5222-5225.
  27. Mues, G. I., T. Z. Munn, and J. D. Raese. 1986. A human gene family with sequence homology to Drosophila melanogaster Hsp70 heat shock gene. J. Biol. Chem. 26:874-877.
  28. Munro, S., and H. R. B. Pelhaim. 1986. An Hsp7O-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein. Cell 46:291-300.
  29. O'Farrell, P. H. 1975. High resolution two-dimensional electro- phoresis of proteins. J. Biol. Chem. 250:4007-4021.
  30. Palter, K. B., M. Watanabe, L. Stinson, A. P. Mahowald, and E. A. Craig. 1986. Expression and localization of Drosophila melanogaster hsp70 cognate proteins. Mol. Cell. Biol. 6:1187- 1203.
  31. Rothstein, R. J. 1983. One step gene disruption in yeast. Methods Enzymol. 101:202-211.
  32. Sherman, F., G. R. Fink, and J. B. Hicks. 1982. Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
  33. Shin, D.-Y., K. Matsumoto, H. lida, I. Uno, and T. Isikawa. 1987. Heat shock response of Saccharomyces cerevisiae mu- tants altered in cyclic AMP-dependent protein phosphorylation. Mol. Cell. Biol. 7:244-250.
  34. Southern, E. M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98:503-517.
  35. Ungewickell, E. 1985. The 70-kd mammalian heat shock proteins are structurally and functionally related to the uncoating protein that releases clathrin triskelia from coated vesicles. EMBO J. 4:3385-3391.
  36. Velasquez, J. M., and S. Lindquist. 1984. Hsp7O: nuclear con- centration during environmental stress and cytoplasmic storage during recovery. Cell 36:655-662.
  37. Welch, W. J., and J. R. Feramisco. 1984. Nuclear and nucleolar localization of the 72,000-dalton heat shock protein in heat- shocked mammalian cells. J. Biol. Chem. 259:4501-4513.