A Novel Cys2His2 Zinc Finger Homolog of AZF1 Modulates Holocellulase Expression in Trichoderma reesei

Author(s):Show less -

Campos Antonieto, Amanda Cristina ^[1] ; Vieira Nogueira, Karoline Maria ^[1] ; de Paula, Renato Graciano ^[1] ; Nora, Lufsa Czamanski ^[1] ; Anzolini Cassiano, Murilo Henrique ^[1] ; Guazzaroni, Maria-Eugenia ^[2] ; Almeida, Fausto ^[1] ; da Silva, Thiago Aparecido ^[1] ; Ries, Laure Nicolas Annick ^[3] ; de Assis, Leandro Jose ^[3] ; Goldman, Gustavo Henrique ^{[3, 4]} ; Silva, Roberto Nascimento ^[1] ; Silva-Rocha, Rafael ^[1]

Affiliation

^[1] Univ Sao Paulo, FMRP, Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, FFCLRP, Ribeirao Preto, SP - Brazil

^[3] Univ Sao Paulo, FCFRP, Ribeirao Preto, SP - Brazil

^[4] Tech Univ Munich, Inst Adv Study, Garching - Germany

• Document type: Journal article
• Source: MSYSTEMS; v. 4, n. 4 JUL-AUG 2019.
• Web of Science Citations: 1

Abstract

Filamentous fungi are remarkable producers of enzymes dedicated to the degradation of sugar polymers found in the plant cell wall. Here, we integrated transcriptomic data to identify novel transcription factors (TFs) related to the control of gene expression of lignocellulosic hydrolases in Trichoderma reesei and Aspergillus nidulans. Using various sets of differentially expressed genes, we identified some putative cis-regulatory elements that were related to known binding sites for Saccharomyces cerevisiae TFs. Comparative genomics allowed the identification of six transcriptional factors in filamentous fungi that have corresponding S. cerevisiae homologs. Additionally, a knockout strain of T. reesei lacking one of these TFs (S. cerevisiae AZF1 homolog) displayed strong reductions in the levels of expression of several cellulase-encoding genes in response to both Avicel and sugarcane bagasse, revealing a new player in the complex regulatory network operating in filamentous fungi during plant biomass degradation. Finally, RNA sequencing (RNA-seq) analysis showed the scope of the AZF1 homologue in regulating a number of processes in T. reesei, and chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) provided evidence for the direct interaction of this TF in the promoter regions of ce17a, ce145a, and swo. Therefore, we identified here a novel TF which plays a positive effect in the expression of cellulase-encoding genes in T. reesei. IMPORTANCE In this work, we used a systems biology approach to map new regulatory interactions in Trichoderma reesei controlling the expression of genes encoding cellulase and hemicellulase. By integrating transcriptomics related to complex biomass degradation, we were able to identify a novel transcriptional regulator which is able to activate the expression of these genes in response to two different cellulose sources. in vivo experimental validation confirmed the role of this new regulator in several other processes related to carbon source utilization and nutrient transport. Therefore, this work revealed novel forms of regulatory interaction in this model system for plant biomass deconstruction and also represented a new approach that could be easy applied to other organisms. (AU)

• FAPESP's process: 16/03322-7 - Role of galectin-3 in the Cryptococcus neoformans infection
• Grantee: Fausto Bruno dos Reis Almeida
• Support type: Research Grants - Young Investigators Grants
• FAPESP's process: 18/03766-8 - Uncovering the regulatory network of Trichoderma reesei: characterization of the RME1 homologue binding motif and identification of new regulators involved in the biomass degradation
• Grantee: Amanda Cristina Campos Antoniêto
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 16/23233-9 - Characterization of calcium-dependent signaling pathway in the secretion and cellulase expression by Trichoderma reesei (Hypocrea jecorina
• Grantee: Renato Graciano de Paula
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 16/03763-3 - Unveiling the mechanisms of gene regulation at the single cell level in Trichoderma reesei
• Grantee: Luísa Czamanski Nora
• Support type: Scholarships in Brazil - Master
• FAPESP's process: 15/04309-1 - Novel approaches to improve functional screening of biocatalysts in metagenomic libraries
• Grantee: María Eugenia Guazzaroni
• Support type: Research Grants - Young Investigators Grants
• FAPESP's process: 12/22921-8 - Synthetic biology approaches for deciphering the logic of signal integration in complex bacterial promoters
• Grantee: Rafael Silva Rocha
• Support type: Research Grants - Young Investigators Grants
• FAPESP's process: 17/04217-5 - Development of a computational tool for searching and strength prediction of bacterial promoters for applications in synthetic biology
• Grantee: Murilo Henrique Anzolini Cassiano
• Support type: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 15/09553-8 - Characterization of new sugar transporters involved in the regulation of lignocellulosic biomass degradation of in Trichoderma reesei
• Grantee: Karoline Maria Vieira Nogueira
• Support type: Scholarships in Brazil - Doctorate