Current Genetics

, Volume 23, Issue 2, pp 115–122

Characterization of the Trichoderma reesei cbh2 promoter

  • Herbert Stangl
  • Franz Gruber
  • Christian P. Kubicek
Original Articles

Abstract

A 613-bp fragment of the 5′ upstream region of the Trichoderma reesei cbh2 gene (coding for the cellulolytic enzyme cellobiohydrolase II) has been isolated and sequenced. Fusion of this fragment to the E. coli uidA gene (coding for β-glucuronidase) leads to-albeit low-expression of β-glucuronidase activity in the presence of cellulose and upon the addition of low molecular weight inducers (sophorose, lactose) of cellobiohydrolase II. It also governed the formation of β-glucuronidase activity during sporulation and its transport to the conidial surface. However, despite the presence of a signal peptide in the cbh2:uidA fusion, β-glucuronidase was not secreted in T. reesei. Defined fragments of the 613-bp promoter region were isolated and used to identify areas involved in the regulation of cbh2 expression by protein-DNA binding assays. At least two binding areas-between-443/-363 and-363/-173, respectively-were identified. In both areas, the DNA-protein complex observed was appreciably larger when cell-free extracts from sophorose-induced mycelia were used. This suggests that at least one of the proteins regulating cbh2 transcription is itself induced by cellulose.

Key words

DNA-protein interactions Trichoderma reseei cbh2 promoter β-glucuronidase 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Berman J, Eisenberg S, Tye BK (1987) Methods Enzymol 155:528–537Google Scholar
  2. Bevan M, Barnes WM, Chilton MD (1983) Nucleic Acids Res 11:369–385Google Scholar
  3. Bram RJ, Kornberg RD (1985) Proc Natl Acad Sci USA 82:43–47Google Scholar
  4. Bisaria VS, Mishra S (1989) CRC Crit Rev Biotechnol 9:61–103Google Scholar
  5. Bunkers GJ (1991) Appl Environ Microbiol 57:2896–2900Google Scholar
  6. Chen CM, Gritzali M, Stafford DW (1987) Bio/Technol 5:274–278Google Scholar
  7. Casabadan MJ, Martinez-Arias A, Shapira SK, Chou J (1983) Methods Enzymol 100:293–308Google Scholar
  8. Davis MA, Hynes MJ (1989) Trends Genet 5:14–19Google Scholar
  9. El-Gogary S, Leite A, Crivellaro O, Eveleight DE, El Dorry H (1989) Proc Natl Acad Sci USA (86:6138–6142Google Scholar
  10. Enari TM, Niku-Paavola ML (1987) CRC Crit Rev Biotechnol 5:67–87Google Scholar
  11. Feinberg AP, Vogelstein B (1983) Analyt Biochem 132:6–13Google Scholar
  12. Fritscher C, Messner R, Kubicek CP (1990) Exp Mycol 14:451–461Google Scholar
  13. Garner MM, Revzin A (1981) Nucleic Acids Res 9:3047–3060Google Scholar
  14. Garoff H (1985) Annu Rev Cell Biol 1:403–445Google Scholar
  15. Gruber F, Visser J, Kubicek CP, De Graaff L (1990a) Curr Genet 18:71–76Google Scholar
  16. Gruber F, Visser J, Kubicek CP, De Graaff L (1990b) Curr Genet 18:447–451Google Scholar
  17. Hager KM, Yanovsky C (1989) Gene 96:153–159Google Scholar
  18. Hamer JE, Timberlake WE (1987) Mol Cell Biol 7:2352–2359Google Scholar
  19. Iturriaga G, Jefferson RA, Bevan MW (1989) Plant Cell 1:381–390Google Scholar
  20. Jefferson RA, Burgess SM, Hirsch D (1986) Proc Natl Acad Sci USA 83:8447–8451Google Scholar
  21. Johnson PF, Knight SL (1989) Annu Rev Biochem 58:799–839Google Scholar
  22. Klionsky DJ, Banta LM, Emr SD (1988) Mol Cell Biol 8:2105–2116Google Scholar
  23. Kolbe J, Kubicek CP (1990) Appl Microbiol Biochenol 34:26–30Google Scholar
  24. Kubicek CP (1987) J Gen Microbiol 133:1481–1487Google Scholar
  25. Kubicek CP, Mühlbauer G, Krotz M, John E, Kubicek-Pranz EM (1988) J Gen Microbiol 134:1215–1222Google Scholar
  26. Kubicek CP, Messner R, Fritscher C, Strauss J, Kubicek-Pranz EM (1990) in: Kubicek CP, Eveleigh DE, Esterbauer H, Steiner W, Kubicek-Pranz EM (eds) Trichoderma cellulases: biochemistry, genetics, physiology and application. Royal Chem Soc, Cambridge UK, pp 101–119Google Scholar
  27. Laemmli UK (1970) Nature 227:680–685Google Scholar
  28. Messner R, Kubicek-Pranz EM, Gsur A, Kubicek CP (1991) Arch Microbiol 155:601–607Google Scholar
  29. Morawetz R, Gruber F, Messner R, Kubicek CP (1992) Curr Genet 21:31–36Google Scholar
  30. Prestridge DS (1991) CABIOS 7:203–206Google Scholar
  31. Punt PJ, Zegers ND, Busscher M, Pouwels PH, van den Hondel CAMJJ (1991) J Biotechnol 17:19–34Google Scholar
  32. Ruiter-Jacobs YMJT de, Broikhuisen M, Unkles S, Campbell EI, Kinghorn JR, Contreras R, Pouwels PH, van den Hondel CAMJJ (1989) Curr Genet 16:159–163Google Scholar
  33. Roberts IN, Oliver RP, Punt PJ, van den Hondel CAMJJ (1989) Curr Genet 15:177–180Google Scholar
  34. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  35. Sanger F, Nicklen S, Coulson AR (1977) Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  36. Seiboth B, Messner R, Gruber F, Kubicek CP (1992) J Gen Microbiol 138:1259–1264Google Scholar
  37. Sternberg D, Mandels G (1979) J Bacteriol 139:761–767Google Scholar
  38. Struhl K (1989) Annu Rev Biochem 58:1051–1077Google Scholar
  39. Tada S, Gomi K, Kitamoto K, Takahashi K, Tamura G, Hara S (1991) Mol Gen Genet 229:301–306Google Scholar
  40. Teeri TT, Lehtovaara P, Kauppinen S, Salovuori I, Knowles J (1987) Gene 51:43–52Google Scholar
  41. Umek RM, Friedman AD, McKnight SL (1991) Science 251:288–292Google Scholar
  42. Vaheri M, Leisola M, Kaupinnen V (1979) Biotechn Lett 1:41–46Google Scholar
  43. Van Gorcom RFM, Pouwels PH, Goosen T, Visser J van den Broek WJ, Hamer JE, Timberlake WE, van den Hondel CAMJJ (1985) Gene 40:99–106Google Scholar
  44. Vanhanen S, Saloheimo A, Ilmen M, Knowles JKC, Penttilä M (1991) Gene 106:129–133Google Scholar
  45. Yanisch-Perron C, Vieira J, Messing J (1985) Gene 33:103–119Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Herbert Stangl
    • 1
  • Franz Gruber
    • 1
  • Christian P. Kubicek
    • 1
  1. 1.Abteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und MikrobiologieTU WienWienAustria

Personalised recommendations