Abstract

The stringent regulatory network of cellulase gene expression in the filamentous fungus Trichoderma reesei involves multiple transcriptional regulators. However, identification and mechanistic investigation of these regulators are still insufficient. Here, we identified a novel transcriptional regulator, CLP1, a plant homeo domain (PHD) Protein that participates in regulating T. reesei cellulase gene expression. Phylogenetic analyses demonstrated that CLP1 homologs are widely distributed in filamentous fungi including Trichoderma, Penicillium, Fusarium, Neurospora, and Aspergillus species. We demonstrated that CLP1 is a nuclear protein and lack of CLP1 significantly impaired the induced expression of cellulase genes. ChIP experiments showed CLP1 binding to the cellulase gene promoters specifically under cellulose conditions and compromised XYR1 occupancy on the same promoters in the absence of CLP1 at the early induction stage. XYR1 overexpression fully rescued the defect in cellulase production but not the defect in conidia formation in the clp1 null mutant. Further analysis showed that the PHD is required for the CLP1 appropriate subcellular localization as well as the induced cellulase gene expression and conidiation. Taken together, these data demonstrated an important role of CLP1 in the regulation of cellulase and xylanase gene expression in T. reesei.

Highlights

  • Rapid and precise regulation of gene expression at the transcriptional level is essential for eukaryotic cells when confronted with environmental stresses and nutrition shift (Znameroski and Glass, 2013; Crawford and Pavitt, 2019)

  • To analyze whether CTI6 homologous protein is present in the filamentous fungus T. reesei, we searched the T. reesei genome with CTI6 as a query and retrieved one gene (Trire2_27020 or TrireRUTC30_1_86967, hereafter named clp1 for CTI6-like protein encoding gene)

  • XYR1 overexpression, had hardly any effect on the compromised conidia formation in Δclp1 (Figures 6E,F). These results suggest that CLP1 participates in modulating cellulase gene expression by facilitating either xyr1 expression or XYR1 binding to cellulase gene promoters, and may contribute to the transcriptional activation process mediated by XYR1

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Summary

Introduction

Rapid and precise regulation of gene expression at the transcriptional level is essential for eukaryotic cells when confronted with environmental stresses and nutrition shift (Znameroski and Glass, 2013; Crawford and Pavitt, 2019). T. reesei has been established as an excellent model system for the investigation of mechanisms underlying induced gene expression (Druzhinina and Kubicek, 2017; Schmoll, 2018). Systematic characterization of the involved transcriptional factors would definitely contribute to understanding the whole regulatory network controlling cellulase gene expression in T. reesei. Eukaryotic gene regulatory network involves multiple transcription factors and cofactors to integrate various environmental signals, and to exert regulatory effect on target genes by acting on their promoters (Tani et al, 2014).

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