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Truncation of the transcriptional repressor protein Cre1 in Trichoderma reesei Rut-C30 turns it into an activator.
Fungal Biol Biotechnol. 2018; 5:15.FB

Abstract

Background

The filamentous fungus Trichoderma reesei (T. reesei) is a natural producer of cellulolytic and xylanolytic enzymes and is therefore industrially used. Many industries require high amounts of enzymes, in particular cellulases. Strain improvement strategies by random mutagenesis yielded the industrial ancestor strain Rut-C30. A key property of Rut-C30 is the partial release from carbon catabolite repression caused by a truncation of the repressor Cre1 (Cre1-96). In the T. reesei wild-type strain a full cre1 deletion leads to pleiotropic effects and strong growth impairment, while the truncated cre1-96 enhances cellulolytic activity without the effect of growth deficiencies. However, it is still unclear which function Cre1-96 has in Rut-C30.

Results

In this study, we deleted and constitutively expressed cre1-96 in Rut-C30. We found that the presence of Cre1-96 in Rut-C30 is crucial for its cellulolytic and xylanolytic performance under inducing conditions. In the case of the constitutively expressed Cre1-96, the cellulase activity could further be improved approximately twofold. The deletion of cre1-96 led to growth deficiencies and morphological abnormalities. An in silico domain prediction revealed that Cre1-96 has all necessary properties that a classic transactivator needs. Consequently, we investigated the cellular localization of Cre1-96 by fluorescence microscopy using an eYFP-tag. Cre1-96 is localized in the fungal nuclei under both, inducing and repressing conditions. Furthermore, chromatin immunoprecipitation revealed an enrichment of Cre1-96 in the upstream regulatory region of the main transactivator of cellulases and xylanases, Xyr1. Interestingly, transcript levels of cre1-96 show the same patterns as the ones of xyr1 under inducing conditions.

Conclusions

The findings suggest that the truncation turns Cre1 into an activating regulator, which primarily exerts its role by approaching the upstream regulatory region of xyr1. The conversion of repressor proteins to potential activators in other biotechnologically used filamentous fungi can be applied to increase their enzyme production capacities.

Authors+Show Affiliations

1Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Gumpendorfer Str. 1a, 1060 Vienna, Austria.2Fungal Genetics and Genomics Lab, Department of Applied Genetics and Cell Biology, BOKU-University of Natural Resources and Life Sciences, Konrad Lorenz Str. 24, 3430 Tulln/Donau, Austria. 3Institute of Microbiology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.2Fungal Genetics and Genomics Lab, Department of Applied Genetics and Cell Biology, BOKU-University of Natural Resources and Life Sciences, Konrad Lorenz Str. 24, 3430 Tulln/Donau, Austria.1Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Gumpendorfer Str. 1a, 1060 Vienna, Austria.1Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Gumpendorfer Str. 1a, 1060 Vienna, Austria.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30151221

Citation

Rassinger, Alice, et al. "Truncation of the Transcriptional Repressor Protein Cre1 in Trichoderma Reesei Rut-C30 Turns It Into an Activator." Fungal Biology and Biotechnology, vol. 5, 2018, p. 15.
Rassinger A, Gacek-Matthews A, Strauss J, et al. Truncation of the transcriptional repressor protein Cre1 in Trichoderma reesei Rut-C30 turns it into an activator. Fungal Biol Biotechnol. 2018;5:15.
Rassinger, A., Gacek-Matthews, A., Strauss, J., Mach, R. L., & Mach-Aigner, A. R. (2018). Truncation of the transcriptional repressor protein Cre1 in Trichoderma reesei Rut-C30 turns it into an activator. Fungal Biology and Biotechnology, 5, 15. https://doi.org/10.1186/s40694-018-0059-0
Rassinger A, et al. Truncation of the Transcriptional Repressor Protein Cre1 in Trichoderma Reesei Rut-C30 Turns It Into an Activator. Fungal Biol Biotechnol. 2018;5:15. PubMed PMID: 30151221.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Truncation of the transcriptional repressor protein Cre1 in Trichoderma reesei Rut-C30 turns it into an activator. AU - Rassinger,Alice, AU - Gacek-Matthews,Agnieszka, AU - Strauss,Joseph, AU - Mach,Robert L, AU - Mach-Aigner,Astrid R, Y1 - 2018/08/20/ PY - 2018/04/15/received PY - 2018/08/02/accepted PY - 2018/8/29/entrez PY - 2018/8/29/pubmed PY - 2018/8/29/medline KW - Carbon catabolite repression KW - Cellulases KW - Chromatin KW - Cre1 KW - Gene regulation KW - Transcription factor KW - Trichoderma reesei KW - Xylanases SP - 15 EP - 15 JF - Fungal biology and biotechnology JO - Fungal Biol Biotechnol VL - 5 N2 - Background: The filamentous fungus Trichoderma reesei (T. reesei) is a natural producer of cellulolytic and xylanolytic enzymes and is therefore industrially used. Many industries require high amounts of enzymes, in particular cellulases. Strain improvement strategies by random mutagenesis yielded the industrial ancestor strain Rut-C30. A key property of Rut-C30 is the partial release from carbon catabolite repression caused by a truncation of the repressor Cre1 (Cre1-96). In the T. reesei wild-type strain a full cre1 deletion leads to pleiotropic effects and strong growth impairment, while the truncated cre1-96 enhances cellulolytic activity without the effect of growth deficiencies. However, it is still unclear which function Cre1-96 has in Rut-C30. Results: In this study, we deleted and constitutively expressed cre1-96 in Rut-C30. We found that the presence of Cre1-96 in Rut-C30 is crucial for its cellulolytic and xylanolytic performance under inducing conditions. In the case of the constitutively expressed Cre1-96, the cellulase activity could further be improved approximately twofold. The deletion of cre1-96 led to growth deficiencies and morphological abnormalities. An in silico domain prediction revealed that Cre1-96 has all necessary properties that a classic transactivator needs. Consequently, we investigated the cellular localization of Cre1-96 by fluorescence microscopy using an eYFP-tag. Cre1-96 is localized in the fungal nuclei under both, inducing and repressing conditions. Furthermore, chromatin immunoprecipitation revealed an enrichment of Cre1-96 in the upstream regulatory region of the main transactivator of cellulases and xylanases, Xyr1. Interestingly, transcript levels of cre1-96 show the same patterns as the ones of xyr1 under inducing conditions. Conclusions: The findings suggest that the truncation turns Cre1 into an activating regulator, which primarily exerts its role by approaching the upstream regulatory region of xyr1. The conversion of repressor proteins to potential activators in other biotechnologically used filamentous fungi can be applied to increase their enzyme production capacities. SN - 2054-3085 UR - https://www.unboundmedicine.com/medline/citation/30151221/Truncation_of_the_transcriptional_repressor_protein_Cre1_in_Trichoderma_reesei_Rut_C30_turns_it_into_an_activator_ L2 - https://fungalbiolbiotech.biomedcentral.com/articles/10.1186/s40694-018-0059-0 DB - PRIME DP - Unbound Medicine ER -
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