Abstract
Basic features of regulation of expression of the genes encoding the cellulases of the filamentous fungus Trichoderma reesei QM9414, the genes cbh1 and cbh2 encoding cellobiohydrolases and the genes egl1, egl2 and egl5 encoding endoglucanases, were studied at the mRNA level. The cellulase genes were coordinately expressed under all conditions studied, with the steady-state mRNA levels of cbh1 being the highest. Solka floc cellulose and the disaccharide sophorose induced expression to almost the same level. Moderate expression was observed when cellobiose or lactose was used as the carbon source. It was found that glycerol and sorbitol do not promote expression but, unlike glucose, do not inhibit it either, because the addition of 1 to 2 mM sophorose to glycerol or sorbitol cultures provokes high cellulase expression levels. These carbon sources thus provide a useful means to study cellulase regulation without significantly affecting the growth of the fungus. RNA slot blot experiments showed that no expression could be observed on glucose-containing medium and that high glucose levels abolish the inducing effect of sophorose. The results clearly show that distinct and clear-cut mechanisms of induction and glucose repression regulate cellulase expression in an actively growing fungus. However, derepression of cellulase expression occurs without apparent addition of an inducer once glucose has been depleted from the medium. This expression seems not to arise simply from starvation, since the lack of carbon or nitrogen as such is not sufficient to trigger significant expression.
Full Text
The Full Text of this article is available as a PDF (1.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abrahão-Neto J., Rossini C. H., el-Gogary S., Henrique-Silva F., Crivellaro O., el-Dorry H. Mitochondrial functions mediate cellulase gene expression in Trichoderma reesei. Biochemistry. 1995 Aug 22;34(33):10456–10462. doi: 10.1021/bi00033a018. [DOI] [PubMed] [Google Scholar]
- Barnett C. C., Berka R. M., Fowler T. Cloning and amplification of the gene encoding an extracellular beta-glucosidase from Trichoderma reesei: evidence for improved rates of saccharification of cellulosic substrates. Biotechnology (N Y) 1991 Jun;9(6):562–567. doi: 10.1038/nbt0691-562. [DOI] [PubMed] [Google Scholar]
- Biely P., Vrsanská M., Claeyssens M. The endo-1,4-beta-glucanase I from Trichoderma reesei. Action on beta-1,4-oligomers and polymers derived from D-glucose and D-xylose. Eur J Biochem. 1991 Aug 15;200(1):157–163. doi: 10.1111/j.1432-1033.1991.tb21062.x. [DOI] [PubMed] [Google Scholar]
- Bisaria V. S., Mishra S. Regulatory aspects of cellulase biosynthesis and secretion. Crit Rev Biotechnol. 1989;9(2):61–103. doi: 10.3109/07388558909040616. [DOI] [PubMed] [Google Scholar]
- Chen H., Hayn M., Esterbauer H. Purification and characterization of two extracellular beta-glucosidases from Trichoderma reesei. Biochim Biophys Acta. 1992 May 22;1121(1-2):54–60. doi: 10.1016/0167-4838(92)90336-c. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Claeyssens M., van Tilbeurgh H., Kamerling J. P., Berg J., Vrsanska M., Biely P. Studies of the cellulolytic system of the filamentous fungus Trichoderma reesei QM 9414. Substrate specificity and transfer activity of endoglucanase I. Biochem J. 1990 Aug 15;270(1):251–256. doi: 10.1042/bj2700251. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark J. T., Kalra P. S., Crowley W. R., Kalra S. P. Neuropeptide Y and human pancreatic polypeptide stimulate feeding behavior in rats. Endocrinology. 1984 Jul;115(1):427–429. doi: 10.1210/endo-115-1-427. [DOI] [PubMed] [Google Scholar]
- Fowler T., Brown R. D., Jr The bgl1 gene encoding extracellular beta-glucosidase from Trichoderma reesei is required for rapid induction of the cellulase complex. Mol Microbiol. 1992 Nov;6(21):3225–3235. doi: 10.1111/j.1365-2958.1992.tb01777.x. [DOI] [PubMed] [Google Scholar]
- Ilmén M., Onnela M. L., Klemsdal S., Keränen S., Penttilä M. Functional analysis of the cellobiohydrolase I promoter of the filamentous fungus Trichoderma reesei. Mol Gen Genet. 1996 Dec 13;253(3):303–314. doi: 10.1007/pl00008597. [DOI] [PubMed] [Google Scholar]
- Ilmén M., Thrane C., Penttilä M. The glucose repressor gene cre1 of Trichoderma: isolation and expression of a full-length and a truncated mutant form. Mol Gen Genet. 1996 Jun 24;251(4):451–460. doi: 10.1007/BF02172374. [DOI] [PubMed] [Google Scholar]
- Kubicek C. P. Involvement of a conidial endoglucanase and a plasma-membrane-bound beta-glucosidase in the induction of endoglucanase synthesis by cellulose in Trichoderma reesei. J Gen Microbiol. 1987 Jun;133(6):1481–1487. doi: 10.1099/00221287-133-6-1481. [DOI] [PubMed] [Google Scholar]
- Kubicek C. P., Messner R., Gruber F., Mach R. L., Kubicek-Pranz E. M. The Trichoderma cellulase regulatory puzzle: from the interior life of a secretory fungus. Enzyme Microb Technol. 1993 Feb;15(2):90–99. doi: 10.1016/0141-0229(93)90030-6. [DOI] [PubMed] [Google Scholar]
- Kulmburg P., Mathieu M., Dowzer C., Kelly J., Felenbok B. Specific binding sites in the alcR and alcA promoters of the ethanol regulon for the CREA repressor mediating carbon catabolite repression in Aspergillus nidulans. Mol Microbiol. 1993 Mar;7(6):847–857. doi: 10.1111/j.1365-2958.1993.tb01175.x. [DOI] [PubMed] [Google Scholar]
- MANDELS M., PARRISH F. W., REESE E. T. Sophorose as an inducer of cellulase in Trichoderma viride. J Bacteriol. 1962 Feb;83:400–408. doi: 10.1128/jb.83.2.400-408.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MANDELS M., REESE E. T. Induction of cellulase in fungi by cellobiose. J Bacteriol. 1960 Jun;79:816–826. doi: 10.1128/jb.79.6.816-826.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mach R. L., Seiboth B., Myasnikov A., Gonzalez R., Strauss J., Harkki A. M., Kubicek C. P. The bgl1 gene of Trichoderma reesei QM 9414 encodes an extracellular, cellulose-inducible beta-glucosidase involved in cellulase induction by sophorose. Mol Microbiol. 1995 May;16(4):687–697. doi: 10.1111/j.1365-2958.1995.tb02430.x. [DOI] [PubMed] [Google Scholar]
- Mandels M., Weber J., Parizek R. Enhanced cellulase production by a mutant of Trichoderma viride. Appl Microbiol. 1971 Jan;21(1):152–154. doi: 10.1128/am.21.1.152-154.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matheucci E., Jr, Henrique-Silva F., el-Gogary S., Rossini C. H., Leite A., Vera J. E., Urioste J. C., Crivellaro O., el-Dorry H. Structure, organization and promoter expression of the actin-encoding gene in Trichoderma reesei. Gene. 1995 Aug 8;161(1):103–106. doi: 10.1016/0378-1119(95)00283-c. [DOI] [PubMed] [Google Scholar]
- Messner R., Kubicek-Pranz E. M., Gsur A., Kubicek C. P. Cellobiohydrolase II is the main conidial-bound cellulase in Trichoderma reesei and other Trichoderma strains. Arch Microbiol. 1991;155(6):601–606. doi: 10.1007/BF00245356. [DOI] [PubMed] [Google Scholar]
- Messner R., Kubicek C. P. Carbon Source Control of Cellobiohydrolase I and II Formation by Trichoderma reesei. Appl Environ Microbiol. 1991 Mar;57(3):630–635. doi: 10.1128/aem.57.3.630-635.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montenecourt B. S., Eveleigh D. E. Preparation of mutants of Trichoderma reesei with enhanced cellulase production. Appl Environ Microbiol. 1977 Dec;34(6):777–782. doi: 10.1128/aem.34.6.777-782.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nehlin J. O., Ronne H. Yeast MIG1 repressor is related to the mammalian early growth response and Wilms' tumour finger proteins. EMBO J. 1990 Sep;9(9):2891–2898. doi: 10.1002/j.1460-2075.1990.tb07479.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nevalainen K. M., Palva E. T. Production of extracellular enzymes in mutants isolated from trichoderma viride unable to hydrolyze cellulose. Appl Environ Microbiol. 1978 Jan;35(1):11–16. doi: 10.1128/aem.35.1.11-16.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nisizawa T., Suzuki H., Nakayama M., Nisizawa K. Inductive formation of cellulase by sophorose in Trichoderma viride. J Biochem. 1971 Sep;70(3):375–385. doi: 10.1093/oxfordjournals.jbchem.a129652. [DOI] [PubMed] [Google Scholar]
- Penttilä M. E., André L., Lehtovaara P., Bailey M., Teeri T. T., Knowles J. K. Efficient secretion of two fungal cellobiohydrolases by Saccharomyces cerevisiae. Gene. 1988;63(1):103–112. doi: 10.1016/0378-1119(88)90549-5. [DOI] [PubMed] [Google Scholar]
- Penttilä M. E., André L., Saloheimo M., Lehtovaara P., Knowles J. K. Expression of two Trichoderma reesei endoglucanases in the yeast Saccharomyces cerevisiae. Yeast. 1987 Sep;3(3):175–185. doi: 10.1002/yea.320030305. [DOI] [PubMed] [Google Scholar]
- Penttilä M., Lehtovaara P., Nevalainen H., Bhikhabhai R., Knowles J. Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene. Gene. 1986;45(3):253–263. doi: 10.1016/0378-1119(86)90023-5. [DOI] [PubMed] [Google Scholar]
- Royer John C., Nakas J. P. Interrelationship of Xylanase Induction and Cellulase Induction of Trichoderma longibrachiatum. Appl Environ Microbiol. 1990 Aug;56(8):2535–2539. doi: 10.1128/aem.56.8.2535-2539.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saloheimo A., Henrissat B., Hoffrén A. M., Teleman O., Penttilä M. A novel, small endoglucanase gene, egl5, from Trichoderma reesei isolated by expression in yeast. Mol Microbiol. 1994 Jul;13(2):219–228. doi: 10.1111/j.1365-2958.1994.tb00417.x. [DOI] [PubMed] [Google Scholar]
- Saloheimo M., Lehtovaara P., Penttilä M., Teeri T. T., Ståhlberg J., Johansson G., Pettersson G., Claeyssens M., Tomme P., Knowles J. K. EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme. Gene. 1988;63(1):11–22. doi: 10.1016/0378-1119(88)90541-0. [DOI] [PubMed] [Google Scholar]
- Shoemaker S. P., Raymond J. C., Bruner R. Cellulases: diversity amongst improved Trichoderma strains. Basic Life Sci. 1981;18:89–109. doi: 10.1007/978-1-4684-3980-9_7. [DOI] [PubMed] [Google Scholar]
- Sternberg D., Mandels G. R. Induction of cellulolytic enzymes in Trichoderma reesei by sophorose. J Bacteriol. 1979 Sep;139(3):761–769. doi: 10.1128/jb.139.3.761-769.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strauss J., Mach R. L., Zeilinger S., Hartler G., Stöffler G., Wolschek M., Kubicek C. P. Cre1, the carbon catabolite repressor protein from Trichoderma reesei. FEBS Lett. 1995 Nov 27;376(1-2):103–107. doi: 10.1016/0014-5793(95)01255-5. [DOI] [PubMed] [Google Scholar]
- Takashima S., Nakamura A., Iikura H., Masaki H., Uozumi T. Cloning of a gene encoding a putative carbon catabolite repressor from Trichoderma reesei. Biosci Biotechnol Biochem. 1996 Jan;60(1):173–176. doi: 10.1271/bbb.60.173. [DOI] [PubMed] [Google Scholar]
- Teeri T. T., Lehtovaara P., Kauppinen S., Salovuori I., Knowles J. Homologous domains in Trichoderma reesei cellulolytic enzymes: gene sequence and expression of cellobiohydrolase II. Gene. 1987;51(1):43–52. doi: 10.1016/0378-1119(87)90472-0. [DOI] [PubMed] [Google Scholar]
- Teeri T. T., Penttilä M., Keränen S., Nevalainen H., Knowles J. K. Structure, function, and genetics of cellulases. Biotechnology. 1992;21:417–445. doi: 10.1016/b978-0-7506-9115-4.50020-6. [DOI] [PubMed] [Google Scholar]
- Vanhanen S., Penttilä M., Lehtovaara P., Knowles J. Isolation and characterization of the 3-phosphoglycerate kinase gene (pgk) from the filamentous fungus Trichoderma reesei. Curr Genet. 1989 Mar;15(3):181–186. doi: 10.1007/BF00435504. [DOI] [PubMed] [Google Scholar]
- Vanhanen S., Saloheimo A., Ilmén M., Knowles J. K., Penttilä M. Promoter structure and expression of the 3-phosphoglycerate kinase-encoding gene (pgk1) of Trichoderma reesei. Gene. 1991 Sep 30;106(1):129–133. doi: 10.1016/0378-1119(91)90577-x. [DOI] [PubMed] [Google Scholar]
- el-Gogary S., Leite A., Crivellaro O., Eveleigh D. E., el-Dorry H. Mechanism by which cellulose triggers cellobiohydrolase I gene expression in Trichoderma reesei. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6138–6141. doi: 10.1073/pnas.86.16.6138. [DOI] [PMC free article] [PubMed] [Google Scholar]