Uncovering the regulatory network of Trichoderma reesei: characterization of the RME1 homologue binding motif and identification of new regulators involved in the biomass degradation

• Grant number: 18/03766-8
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: August 01, 2018
• End date: December 31, 2021
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Roberto do Nascimento Silva
• Grantee: Amanda Cristina Campos Antoniêto
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The filamentous fungus Trichoderma reesei is known for its high capacity in secrete cellulolytic enzymes that act in the hydrolysis of cellulose, which is a key point in the production of bioethanol. The gene expression of fungal cellulases is finely controlled at transcriptional level according to the available carbon source in the culture medium. Despite the numerous studies on the regulation of cellulolytic gene expression in T. reesei, several transcription factors as well as the mechanisms by which these regulators interact with the cis-regulatory elements in the promoter regions of the cellulolytic genes are still unclear. Thus, this proposal aims to contribute to the elucidation of the regulatory network of T. reesei and is composed of two approaches that are interrelated: first, a new regulator not yet characterized in T. reesei, homologous to the RME1 of Sacchamomyces cerevisiae, will be produced recombinantly in Escherichia coli and characterized regarding to its DNA-binding dynamics by the Surface Plasmon Resonance technology, obtaining parameters of specificity, kinetics and affinity of the molecular interactions. In parallel, the overall identification of the transcription factors that act during conditions of carbon catabolic repression and induction of cellulase production will be performed. For this, the T. reesei strain QM6a will be grown in sugarcane bagasse and the nuclear fraction obtained will be submitted to the Promoter Trapping method, in which the transcription factors that interact with the promoter region of the genes involved with the biomass degradation will be purified and identified by mass spectrometry. The knowledge generated through this study will be crucial to understand the role of the regulators of T. reesei on a global scale, expanding the biotechnological potential of this fungus in the field of biofuels and in several other industrial applications. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CAMPOS ANTONIETO, AMANDA CRISTINA; VIEIRA NOGUEIRA, KAROLINE MARIA; DE PAULA, RENATO GRACIANO; NORA, LUFSA CZAMANSKI; ANZOLINI CASSIANO, MURILO HENRIQUE; GUAZZARONI, MARIA-EUGENIA; ALMEIDA, FAUSTO; DA SILVA, THIAGO APARECIDO; RIES, LAURE NICOLAS ANNICK; DE ASSIS, LEANDRO JOSE; et al. A Novel Cys2His2 Zinc Finger Homolog of AZF1 Modulates Holocellulase Expression in Trichoderma reesei. MSYSTEMS, v. 4, n. 4, JUL-AUG 2019. (16/23233-9, 16/03322-7, 15/04309-1, 17/04217-5, 16/03763-3, 18/03766-8, 15/09553-8, 12/22921-8)

CAMPOS ANTONIETO, AMANDA CRISTINA; MAUES, DAVID BATISTA; VIEIRA NOGUEIRA, KAROLINE MARIA; DE PAULA, RENATO GRACIANO; STEINDORFF, ANDREI STECCA; KENNEDY, JOHN F.; PANDEY, ASHOK; GUPTA, VIJAI KUMAR; SILVA, ROBERTO N.. Engineering of holocellulase in biomass-degrading fungi for sustainable biofuel production. JOURNAL OF CLEANER PRODUCTION, v. 371, p. 13-pg., 2022-10-15. (18/03766-8, 19/11655-4)