1887

Abstract

The constitutive and the heat-shock-induced expression of members of heat-shock protein families changed during vegetative development and conidiation of as determined by two-dimensional gel electrophoresis. Western blot, and ELISA analyses revealed the highest amounts of the constitutive heat-shock protein 70 (HSC70) in conidiating aerial hyphae and dormant conidia. During conidial germination the amount of HSC70 decreased and subsequently increased during vegetative growth. Stationary mycelia and young aerial hyphae exhibited the lowest HSC70 level. The stationary-phase-dependent decrease in HSC70 was accompanied by a concomitant increase in its nuclear localization, whereas no significant changes in the amount of nuclear HSC70 were found during aerial hyphae development. The cAMP content during aerial hyphae development was inversely correlated with that of HSC70. To examine possible causal relations between HSC70 expression and cAMP content, the adenylate-cyclase-deficient mutant (-1) was analysed, which exhibits low concentrations of endogenous cAMP. This mutant, however, showed a lower constitutive HSC70 level, compared to the strain. Treatment of the strain and -1 mutant with 20 μM 8-bromo-cAMP did not result in significant changes of the constitutive HSC70 level, but in the level of heat-induced HSC/HSP70. In a developmental mutant (acon-2) which is defective in a differentiation step toward conidiation, the expression of HSC70 in aerial hyphae was delayed until the first proconidial chains were observed. It is concluded that the differential expression of HSC/HSP70 does not depend on different nuclear levels of HSC70 or on changes in cAMP concentrations, but rather on developmental genes controlling conidiation.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-11-3615
1997-11-01
2024-12-01
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/11/mic-143-11-3615.html?itemId=/content/journal/micro/10.1099/00221287-143-11-3615&mimeType=html&fmt=ahah

References

  1. Becker J., Craig E.A. 1994; Heat-shock proteins as molecular chaperones.. Eur J Biochem 219:11–23
    [Google Scholar]
  2. Bienz M., Pelham H.R.B. 1987; Mechanisms of heat shock activation in higher eukaryotes.. Adv Genet 24:31–72
    [Google Scholar]
  3. Bonato M.C., Silvia A.M., Gomes S.L., Maia J.C., Juliani M.H. 1987; Differential expression of heat shock proteins and spontaneous synthesis of HSP70 during the life cycle of Blasto-cladiella emersonii.. Eur J Biochem 163:211–220
    [Google Scholar]
  4. Bonner W.M., Laskey R.A. 1974; A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels.. Eur J Biochem 46:83–88
    [Google Scholar]
  5. Boorstein W.R., Craig E.A. 1990; Regulation of a yeast HSP70 gene by a cAMP responsive transcriptional control element.. EMBO J 9:2543–2553
    [Google Scholar]
  6. Bruce J.L., Price B.D., Coleman C.N., Calderwood S.K. 1993; Oxidative injury rapidly activates the heat shock transcription factor but fails to increase levels of heat shock proteins.. Cancer Res 53:12–15
    [Google Scholar]
  7. Cameron S., Levin L., Zooler M., Wigler M. 1988; cAMP-independent control of sporulation, glycogen metabolism, and heat shock resistance in Saccharomyces cerevisiae.. Cell 53:555–566
    [Google Scholar]
  8. Choi H.-S., Li B., Lin Z., Huang E., Liu A.Y.-C. 1991; cAMP and cAMP-dependent protein kinase regulate the human heat shock protein 70 gene promoter activity.. J Biol Chem 266:11858–11865
    [Google Scholar]
  9. Coca M.A., Almoguera C., Jordano J. 1994; Expression of sunflower low-molecular-weight heat-shock proteins during embryogenesis and persistence after germination: localization and possible functional implications.. Plant Mol Biol 25:479–492
    [Google Scholar]
  10. Cruz A.K., Terenzi H.F., Jorge J.A., Terenzi H.F. 1988; Cyclic AMP-dependent, constitutive thermotolerance in the adenylate cyclase-deficient cr-1 (crisp) mutant of Neurospora crassa.. Curr Genet 13:451–454
    [Google Scholar]
  11. Dice J.F., Agarraberes F., Kirven-Brooks M., Terlecky L.J., Terlecky S.R. 1994; Heat shock 70-kD proteins and lysosomal proteolysis.. In The Biology of Heat Shock Proteins and Molecular Chaperones pp. 137–152 Edited by Morimoto R. I., Tissieres A., Georgopoulos C. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  12. Engelberg D., Zandi E., Parker C.S., Karin M. 1994; The yeast and mammalian Ras pathway control transcription of heat shock genes independently of heat shock transcription factor.. Mol Cell Biol 14:4929–4937
    [Google Scholar]
  13. Fracella F., Mohsenzadeh S., Rensing L. 1993; Purification and partial amino acid sequence of the major 70,000-Dalton heat shock protein in Neurospora crassa.. Exp My col 17:362–367
    [Google Scholar]
  14. Giardina C., Lis J.T. 1995; Dynamic protein-DNA architecture of a yeast heat shock promoter.. Mol Cell Biol 15:2737–2744
    [Google Scholar]
  15. Hartl F.-U., Hlodan R., Langer T. 1994; Molecular chaperones in protein folding: the art of avoiding sticky situations.. Trends Biochem Sci 19:20–25
    [Google Scholar]
  16. Hensold J.O., Hunt C.R., Calderwood S.K., Housman D.E., Kingston R.E. 1990; DNA binding of heat shock factor to the heat shock element is insufficient for transcriptional activation in murine erythroleukemia cells.. Mol Cell Biol 10:1600–1608
    [Google Scholar]
  17. Hightower L., Nover L. editors 1991 Results and Problems in Cell Differentiation: Heat Shock and Development. Heidelberg: Springer;
    [Google Scholar]
  18. Horowitz N.H. 1947; Methionine synthesis in Neurospora.The isolation of cystathionine.. J Biol Chem 171:255–264
    [Google Scholar]
  19. Iida H., Yahara I. 1984; A heat shock-resistant mutant of Saccharomyces cerevisiae shows constitutive synthesis of two heat shock proteins and altered growth.. J Cell Biol 99:1441–1450
    [Google Scholar]
  20. Jurivich D., Sistonen L., Kroes R., Morimoto R.I. 1992; Effect of sodium salicylate on the human heat shock response.. Science 255:1243–1245
    [Google Scholar]
  21. Kapoor M., Curle C.A., Runham C. 1995; The hsp70 gene family of Neurospora crassa: cloning, sequence analysis, expression, and genetic mapping of the major stress-inducible member.. J Bacteriol 177:212–221
    [Google Scholar]
  22. Kiang J.G., Carr F.E., Burns M.E., McClain D.E. 1994; HSP-72 synthesis is promoted by increase in [Ca2+]i or activation of G proteins but not pHi or cAMP.. Am J Physiol 267:C104–C114
    [Google Scholar]
  23. Kim D., Ouyang H., Li G.C. 1995; Heat shock protein Hsp70 accelerates the recovery of heat-shocked mammalian cells through its modulation of heat shock transcription factor HSF1.. Proc Natl Acad Sci USA 922126–2130
    [Google Scholar]
  24. Kurtz S., Lindquist S. 1984; Changing patterns of gene expression during sporulation in yeast.. Proc Natl Acad Sci USA 817323–7327
    [Google Scholar]
  25. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  26. Liu R.Y., Kim D., Yang S.H., Li G.C. 1993; Dual control of heat shock response: involvement of a constitutive heat shock element-binding factor.. Proc Natl Acad Sci USA 903078–3082
    [Google Scholar]
  27. Loros J.J., Dunlap J.C. 1991; Neurospora crassa clock-controlled genes are regulated at the level of transcription.. Mol Cell Biol 11:558–563
    [Google Scholar]
  28. Machwe A., Kapoor M. 1993; Identification of the heat shock protein of Neurospora crassa corresponding to the stress-inducible peroxidase.. Biochem Biophys Res Commun 196:692–698
    [Google Scholar]
  29. Matsumoto K., Uno I., Ishikawa T. 1983; Initiation of meiosis in yeast mutants defective in adenylate cyclase and cyclic AMP- dependent protein kinase.. Cell 32:417–423
    [Google Scholar]
  30. Matsuyama S.S., Nelson R.E., Siegel R.W. 1974; Mutations specifically blocking differentiation of macroconidia in Neurospora crassa.. Dev Biol 41:278–287
    [Google Scholar]
  31. Moreau N., Lain M.-C., Billoud B., Angelier N. 1994; Transcription of amphibian lampbrush chromosomes is disturbed by microinjection of HSP70 monoclonal antibodies.. Exp Cell Res 211:108–114
    [Google Scholar]
  32. Morimoto R.I., Sarge K.D., Abravaya K. 1992; Transcriptional regulation of heat shock genes.. J Biol Chem 267:21987–21990
    [Google Scholar]
  33. Morimoto R.I., Tissieres A., Georgopoulos C. editors 1994 The Biology of Heat Shock Proteins and Molecular Chaperones. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  34. Nelson R. E., Selitrennikoff C. P., Siegel R. W. . 1975; Cell changes in Neurospora.. In Results and Problems in Cell Differentiation: Cell Cycle and Cell Differentiation pp. 291–310 Edited by Reinert J., Holzer H. Heidelberg: Springer;
    [Google Scholar]
  35. Neuhoff V., Philipp K., Zimmer H. G., Mesecke S. 1979; A simple, versatile, sensitive and volume-independent method for quantitative protein determination which is independent of other external influences.. Hoppe-Seylers Z Physiol Chem 3601657–1670
    [Google Scholar]
  36. Ninnemann H. 1991; Photostimulation of conidiation in mutants of Neurospora crassa.. Photochem Photobiol B-Biol 9:189–199
    [Google Scholar]
  37. O’Farrell P.H. 1975; High resolution two-dimensional electrophoresis of proteins.. J Biol Chem 250:4007–4021
    [Google Scholar]
  38. Patterson N.A., Kapoor M. 1995; Developmental-regulated expression of heat shock genes in Leptosphaeria maculans.. Can J Biochem 41:499–507
    [Google Scholar]
  39. Pizurki L., Polla B. 1994; cAMP modulates stress protein synthesis in human monocytes-macrophages.. J Cell Physiol 161:169–177
    [Google Scholar]
  40. Plesofsky-Vig N., Brambl R. 1985a; The heat shock response of Neurospora crassa: protein synthesis and induced thermotolerance.. J Bacteriol 162:1083–1091
    [Google Scholar]
  41. Plesofsky-Vig N., Brambl R. 1985b; The heat shock response of fungi.. Exp Mycol 9:187–194
    [Google Scholar]
  42. Plesofsky-Vig N., Brambl R. 1990; Gene sequence and analysis of hsp30, a small heat shock protein of Neurospora crassa which associates with mitochondria.. J Biol Chem 265:15432–15440
    [Google Scholar]
  43. Rabindran S.K., Wisniewski J., Li L., Li G.C., Wu C. 1994; Interaction between heat shock factor and Hsp70 is insufficient to suppress induction of DNA-binding activity in vivo.. Mol Cell Biol 14:6552–6560
    [Google Scholar]
  44. Roberts A., Berlin V., Hager K.M., Yanofsky C. 1988; Molecular analysis of a Neurospora crassa gene expressed during conidiation.. Mol Cell Biol 8:2411–2418
    [Google Scholar]
  45. Roychowdhury H.S., Wong D., Kapoor M. 1992; hsp80 of Neurospora crassa: cDNA cloning, gene mapping, and studies of mRNA accumulation under stress.. Biochem Cell Biol 70:1356–1367
    [Google Scholar]
  46. Russo V.E.A., Pandit N.N. 1992; Development in N. crassa.. In Development - the Molecular Genetic Approach pp. 88–102 Edited by Russo V. E. A., Brody S., Cove D., Ottolenghi S. Heidelberg: Springer;
    [Google Scholar]
  47. Shi Y., Thomas J.O. 1992; The transport of proteins into the nucleus requires the 70-kilodalton heat shock protein or its cytosolic cognate.. Mol Cell Biol 12:2186–2192
    [Google Scholar]
  48. Shin D.-Y., Matsumoto K., lida H., Uno I., Ishikawa T. 1987; Heat shock response of Saccharomyces cerevisiae mutants altered in cyclic AMP-dependent protein phosphorylation.. Mol Cell Biol 7:244–250
    [Google Scholar]
  49. Sorger P.K., Pelham H.R.B. 1988; Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation.. Cell 54:855–864
    [Google Scholar]
  50. Sorger P.K., Lewis M.J., Pelham H.R.B. 1987; Heat shock factor is regulated differently in yeast and HeLa cells.. Nature 329:81–84
    [Google Scholar]
  51. Springer M., Yanofsky C. 1989; A morphological and genetic analysis of conidiophore development in Neurospora crassa.. Genes Dev 3:559–571
    [Google Scholar]
  52. Terenzi H., Flawia M., Torres H. 1974; A Neurospora crassa morphological mutant showing reduced adenylate cyclase activity.. Biochem Biophys Res Commun 58:990–996
    [Google Scholar]
  53. Toledo I., Rangel P., Hansberg W. 1995; Redox imbalance at the start of each morphogenetic step of Neurospora crassa conidiation.. Arch Biochem Biophys 319:519–524
    [Google Scholar]
  54. Towbin H., Staehelin T.M., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.. Proc Natl Acad Sci USA 764350–4354
    [Google Scholar]
  55. Vassilev A.O., Plesofsky-Vig N., Brambl R. 1992; Isolation, partial amino acid sequence, and cellular distribution of heat-shock protein Hsp98 from Neurospora crassa.. Biochim Biophys Acta 1156:1–6
    [Google Scholar]
  56. Vogel H.J. 1956; A convenient growth medium for Neurospora (medium M).. Microb Genet Bull 13:42–43
    [Google Scholar]
  57. Wang C., Lin B.L. 1993; The disappearance of an HSC70 species in mung bean seed during germination: purification and characterization of the protein.. Plant Mol Biol 21:317–329
    [Google Scholar]
  58. Werner-Washburne M., Becker J., Kosic-Smithers J., Craig E.A. 1989; Yeast HSP70 RNA levels vary in response to the physiological status of the cell.. J Bacteriol 171:2680–2688
    [Google Scholar]
  59. Williams G.T., McClanahan T.L., Morimoto R.I. 1989; Ela transactivation of the human HSP70 promoter is mediated through the basal transcriptional complex.. Mol Cell Biol 9:2574–2587
    [Google Scholar]
  60. Wu B.J., Kingston R.E., Morimoto R.I. 1987; Detection of three protein binding sites in the serum-regulated promoter of the human gene encoding the 70-kDa heat shock protein.. Proc Natl Acad Sci USA 842203–2207
    [Google Scholar]
/content/journal/micro/10.1099/00221287-143-11-3615
Loading
/content/journal/micro/10.1099/00221287-143-11-3615
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error