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Molecular Genetics and Genomics

, Volume 270, Issue 1, pp 46–55 | Cite as

Nucleosome transactions on the Hypocrea jecorina ( Trichoderma reesei) cellulase promoter cbh2 associated with cellulase induction

  • S. ZeilingerEmail author
  • M. Schmoll
  • M. Pail
  • R. L. Mach
  • C. P. Kubicek
Original Paper

Abstract

The 5′ regulatory region of the cbh2 gene of Hypocrea jecorina contains the cbh2 activating element (CAE) which is essential for induction of cbh2 gene expression by sophorose and cellulose. The CAE consists of two motifs, a CCAAT box on the template strand and a GTAATA box on the coding strand, which cooperate during induction. Northern analyses of cbh2 gene expression has revealed an absolute dependence on induction, but no direct effect of Cre1-mediated carbon catabolite repression. Investigation of the chromatin structure in the wild-type strain showed that, under repressing conditions, there is a nucleosome free region (nfr) around the CAE, which is flanked by strictly positioned nucleosomes. Induction results in a loss of positioning of nucleosomes −1 and −2 downstream of the CAE, thus making the TATA box accessible. Simultaneous mutation of both motifs of the CAE, or of the CCAAT-box alone, also leads to shifting of nucleosome −1, which normally covers the TATA-box under repressing conditions, whereas mutation of the GTAATA element results in a narrowing of the nfr, indicating that the proteins that bind to both motifs in the CAE interact with chromatin, although in different ways. A cellulase-negative mutant strain, which has previously been shown to be altered in protein binding to the CAE, still displayed the induction-specific changes in nucleosome structure, indicating that none of the proteins that directly interact with CAE are affected, and that nucleosome rearrangement and induction of cbh2 expression are uncoupled. Interestingly, the carbon catabolite repressor Cre1 is essential for strict nucleosome positioning in the 5′ regulatory sequences of cbh2 under all of the conditions tested, and induction can occur in a promoter that lacks positioned nucleosomes. These data suggest that Cre1, the Hap2/3/5 complex and the GTAATA-binding protein are all involved in nucleosome assembly on the cbh2 promoter, and that the latter two respond to inducing conditions by repositioning nucleosome −1.

Keywords

Hypocrea jecorina  Trichoderma reesei Chromatin  cbh2 promoter 

Notes

Acknowledgements

This work was supported by a grant from the Fonds zur Förderung Wissenschaftlicher Forschung (P13672) to CPK. We thank A. Czsifersky for providing strain CK11

References

  1. Agalioti T, Lomvardas S, Parekh BS, Maniatis T, Thanos D (2000) Ordered recruitment of chromatin modifying and general transcription factors to the IFN-β promoter. Cell 103:667–678PubMedGoogle Scholar
  2. Aro N, Saloheimo A, Ilmen M, Penttilä M (2001) ACEII, a novel transcriptional activator involved in regulation of cellulase and xylanase genes of Trichoderma reesei. J Biol Chem 276:24309–24314PubMedGoogle Scholar
  3. Beato M, Eisfeld K (1997) Transcription factor access to chromatin. Nucleic Acids Res 25:3559–3563CrossRefPubMedGoogle Scholar
  4. Chomczynski P, Sacchi N (1987) Single step method for RNA isolation by acidic guanidinium thiocyanate-phenol-chlorophorm extraction. Anal Biochem 162:156–159PubMedGoogle Scholar
  5. Cziferszky A, Mach RL, Kubicek CP (2002) Phosphorylation positively regulates DNA-binding of the carbon catabolite repressor Cre1 of Hypocrea jecorina ( Trichoderma reesei). J Biol Chem 277:14688–14694PubMedGoogle Scholar
  6. Deckert J, Struhl K (2001) Histone acetylation at promoters is differentially affected by specific activators and repressors. Mol Cell Biol 21:2726–2735CrossRefPubMedGoogle Scholar
  7. Di Mauro E, Kendrew SG, Caserta M (2000) Two distinct nucleosome alterations characterize chromatin remodeling at the Saccharomyces cerevisiae ADH2 promoter. J Biol Chem 275:7612–7618CrossRefPubMedGoogle Scholar
  8. Dowzer CE, Kelly JM (1991) Analysis of the creA gene, a regulator of carbon catabolite repression in Aspergillus nidulans. Mol Cell Biol 11: 5701–5709PubMedGoogle Scholar
  9. Gonzalez R, Scazzocchio C (1997) A rapid method for chromatin structure analysis in the filamentous fungus Aspergillus nidulans. Nucleic Acids Res 25:3955–3956CrossRefPubMedGoogle Scholar
  10. Gregory PD (2001) Transcription and chromatin converge: lessons from yeast genetics. Curr Opin Genet Dev 11:142–147CrossRefPubMedGoogle Scholar
  11. Gregory PD, Schmid A, Zavari M, Münsterkötter M, Hörz W (1999) Chromatin remodelling at the PHO8 promoter requires SWI-SNF and SAGA at a step subsequent to activator binding. EMBO J 18:6407–6414PubMedGoogle Scholar
  12. Hynes MJ, Davis MA (1996) Regulation of acetamide catabolism. In: Brambl R, Marzluf G (eds) The Mycota III: biochemistry and molecular biology. Springer, New York, pp 381–393Google Scholar
  13. Ilmen M, Thrane C, Penttilä M (1996a) The glucose repressor gene cre1 of Trichoderma: isolation and expression of a full-length and a truncated mutant form. Mol Gen Genet 251:451–460PubMedGoogle Scholar
  14. Ilmen M, Onnela ML, Klemsdal S, Keränen S, Penttilä M (1996b) Functional analysis of the cellobiohydrolase I promoter of the filamentous fungus Trichoderma reesei. Mol Gen Genet 253:303–314PubMedGoogle Scholar
  15. Ilmen M, Saloheimo A, Onnela ML, Penttilä M (1997) Regulation of cellulase gene expression in the filamentous fungus Trichoderma reesei. Appl Environ Microbiol 63:1298–1306PubMedGoogle Scholar
  16. Krebs JE, Fry CJ, Samuels ML, Peterson CL (2000) Global role for chromatin remodeling enzymes in mitotic gene expression. Cell 102:587–598PubMedGoogle Scholar
  17. Kubicek CP, Penttilä M (1998) Regulation of production of plant polysaccharide degrading enzymes by Trichoderma reesei. In: Harman GE, Kubicek CP (eds) Trichoderma and Gliocladium, vol 2. Taylor and Francis, London, pp 49–72Google Scholar
  18. Kubicek CP, Messner R, Gruber F, Mandels M, Kubicek-Pranz EM (1993) Triggering of cellulase biosynthesis by cellulose in Trichoderma reesei. Involvement of a constitutive, sophorose-inducible, glucose-inhibited beta-diglucoside permease. J Biol Chem 268:19364–19368PubMedGoogle Scholar
  19. Mach RL, Schindler M, Kubicek CP (1994) Transformation of Trichoderma reesei based on hygromycinB resistance using homologous expression signals. Curr Genet 25:567–570PubMedGoogle Scholar
  20. Mach RL, Strauss J, Zeilinger S, Schindler M, Kubicek CP (1996) Carbon catabolite repression of xylanase I ( xyn1) gene expression in Trichoderma reesei. Mol Microbiol 21:1273–1281PubMedGoogle Scholar
  21. McPherson CE, Shim EY, Friedman DS, Zaret KS (1993) An active tissue-specific enhancer and bound transcription factors existing in a precisely positioned nucleosomal array. Cell 75:387–398PubMedGoogle Scholar
  22. Merika M, Thanos D (2001) Enhanceosomes. Curr Opin Biochem Mol Biophys 11:205–208CrossRefGoogle Scholar
  23. Montenecourt BS, Eveleigh DE (1979) Selective screening methods for the isolation of high yielding cellulase mutants of Trichoderma reesei. Adv Chem 181: 289–301Google Scholar
  24. Muro-Pastor M, Gonzalez R, Strauss J, Narendja F, Scazzocchio C (1999) The GATA factor AreA is essential for chromatin remodelling in a eukaryotic bidirectional promoter. EMBO J 18:1584–1597PubMedGoogle Scholar
  25. Narendja F, Davis MA, Hynes MJ (1999) AnCF, the CCAAT binding complex of Aspergillus nidulans, is essential for the formation of a DNaseI-hypersensitive site in the 5′ region of the amdS gene. Mol Cell Biol 19:6523–6531PubMedGoogle Scholar
  26. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual (2nd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.Google Scholar
  27. Seiboth B, Messner R, Gruber F, Kubicek CP (1992) Disruption of the Trichoderma reesei cbh2 gene coding for cellobiohydrolase II leads to a delay in the triggering of cellulase formation by cellulose. J Gen Microbiol 138:1259–1264Google Scholar
  28. Shen CH, Leblanc BP, Alfieri JA, Clark DJ (2001) Remodeling of yeast CUP1 chromatin involves activator-dependent repositioning of nucleosomes over the entire gene and flanking sequences. Mol Cell Biol 21:534–547CrossRefPubMedGoogle Scholar
  29. Stangl H, Gruber F, Kubicek CP (1993) Characterization of the Trichoderma reesei cbh2 promoter. Curr Genet 23:115–122PubMedGoogle Scholar
  30. Steidl S, Papagiannopoulos P, Litzka O, Afrianopoulos A, Davis MA, Brakhage A, Hynes MJ (1999) AnCF, the CCAAT binding complex of Aspergillus nidulans, contains products of the hapB, hapC and hapE genes and is required for activation by the pathway-specific regulatory gene amdR. Mol Cell Biol 19:99–106PubMedGoogle Scholar
  31. Strauss J, Mach RL, Zeilinger S, Hartler G, Stöffler G, Wolschek M, Kubicek CP (1995) Cre1, the carbon catabolite repressor protein from Trichoderma reesei. FEBS Lett 376:103–107PubMedGoogle Scholar
  32. Svaren J, Hörz W (1997) Transcription factors vs. nucleosomes: regulation of the PHO5 promoter in yeast. Trends Biochem Sci 22:93–97CrossRefPubMedGoogle Scholar
  33. Tanaka A, Kamei K, Tanoue S, Papagiannopoulos P, Steidl S, Brakhage AA, Davis MA, Hynes MJ, Kato M, Kobayashi T, Tsukagoshi N (2001) AoHapB, AoHapC and AoHapE, subunits of the Aspergillus oryzae CCAAT-binding complex, are functionally interchangable with the corresponding subunits in Aspergillus nidulans. Curr Genet 39:175–182CrossRefPubMedGoogle Scholar
  34. Wolschek MF, Narendja F, Karlseder J, Kubicek CP, Scazzocchio C, Strauss J (1998) In situ detection of protein-DNA interactions in filamentous fungi by in vivo footprinting. Nucleic Acids Res 26:3862–3864PubMedGoogle Scholar
  35. Workman JL, Kingston RE (1998) Alteration of nucleosome structure as a mechanism of transcriptional regulation. Annu Rev Biochem 67:545–579PubMedGoogle Scholar
  36. Wu C (1980) The 5′ ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature 286:854–860PubMedGoogle Scholar
  37. Yu Y, Eriksson P, Stillman DJ (2000) Architectural transcription factors and the SAGA complex function in parallel pathways to activate transcription. Mol Cell Biol 20:2350–2357CrossRefPubMedGoogle Scholar
  38. Zeilinger S, Mach RL, Kubicek CP (1998) Two adjacent protein binding motifs in the cbh2 (cellobiohydrolase II-encoding) promoter of the fungus Hypocrea jecorina ( Trichoderma reesei) cooperate in the induction by cellulose. J Biol Chem 273:34463–34471PubMedGoogle Scholar
  39. Zeilinger S, Haller M, Mach RL, Kubicek CP (2000) Molecular characterization of a cellulase-negative mutant of Hypocrea jecorina. Biochem Biophys Res Comm 277:581–588PubMedGoogle Scholar
  40. Zeilinger S, Ebner A, Marosits T, Mach RL, Kubicek CP (2001) The Hypocrea jecorina Hap 2/3/5 protein complex binds to the inverted CCAAT-box (ATTGG) within the cbh2 (cellobiohydrolase II-gene) activating element. Mol Genet Genomics 266:56–63PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • S. Zeilinger
    • 1
    Email author
  • M. Schmoll
    • 1
  • M. Pail
    • 1
  • R. L. Mach
    • 1
  • C. P. Kubicek
    • 1
  1. 1.Microbial Biochemistry and Gene Technology Department, Institute for Chemical EngineeringTechnical University of ViennaWienAustria

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