The GPI-Anchored GH76 Protein Dfg5 Affects Hyphal Morphology and Osmoregulation in the Mycoparasite Trichoderma atroviride and Is Interconnected With MAPK Signaling

Atanasova, Lea; Moreno-Ruiz, Dubraska; Grünwald‐Gruber, Clemens; Hell, Viktoria; Zeilinger, Susanne

doi:10.3389/fmicb.2021.601113

Cited by 8 publications

(3 citation statements)

References 70 publications

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“…It was confirmed in Saccharomyces cerevisiae confirmed that Hog1 and Slt2 cell signaling pathways are affected in the dfg5Δ mutant [7]. Further evidence in the mycoparasite Trichoderma atroviride, also suggests that Dfg5 plays critical role in hyphal morphogenesis and osmoregulation via MAPK signaling [8] Our past studies in C. albicans have utilized the pMET3 modulated dfg5/dcw1 conditional mutant [6]. We showed that DFG5 and DCW1 function in covalent incorporation of cell wall proteins and thus play critical roles in cell wall biogenesis [9].…”

Section: Introductionmentioning

confidence: 78%

<i>Candida albicans</i> Mannosidases, Dfg5 and Dcw1, Are Required for Pathogenesis

Razmi,

Kim,

Chinnici

et al. 2024

Preprint

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C. albicans Dfg5 and Dcw1 are homologous cell wall alpha-1,6-mannosidases with important functions. Our past studies have shown that Dfg5 and Dcw1 function in cell wall biogenesis via cross-linking of glycoproteins into the cell wall, thus playing a key role in cell wall integrity. Additionally, Dfg5 and Dcw1 are required for hyphal morphogenesis. However, the exact functions of Dfg5 and Dcw1 in cell wall integrity, hyphal morphogenesis, and pathogenesis were not known. In this study, we determined the relation of Dfg5 and Dcw1 with Hog1 MAPK, that plays a key role in cell wall integrity via regulation of chitin synthesis in C. albicans. Additionally, we also determined the effect of dfg5 and dcw1 mutations on gene expression of transcriptional regulators of hyphal morphogenesis. Furthermore, we determined the effects of dfg5 and dcw1 mutations on pathogenesis in a mouse model of oral candidiasis. Our results demonstrate that dfg5 and dcw1 mutations, as well as hog1 knock-out mutation result in dysregulation of chitin synthesis resulting in a cell separation defect. The heterozygous and conditional mutations of dfg5 and dcw1 result in decreased levels of Cst20, a positive regulator of hyphal morphogenesis. However, mutations of dfg5 and dcw1 mutations resulted in increased levels of all the five negative regulators of hyphal morphogenesis – Tup1, Nrg1, Mig1, Rbf1 and Rfg1. However, Tup1 levels were significantly higher than other negative regulators indicating that Dfg5 and Dcw1 function in hyphal morphogenesis by repression of Tup1. Finally, dfg5 and dcw1 mutations affected the ability of C. albicans to cause oral candidiasis in mice. Thus, cell wall glycosidases, Dfg5 and Dcw1, are required for virulence and pathogenesis and represent novel drug targets.

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Section: Introductionmentioning

confidence: 78%

<i>Candida albicans</i> Mannosidases, Dfg5 and Dcw1, Are Required for Pathogenesis

Razmi,

Kim,

Chinnici

et al. 2024

Preprint

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“…Accordingly, the genomes of Trichoderma species encode three types of MAPK. They belong to the so-called pathogenicity Fus3, cell-wall integrity Slt2, and osmoregulatory Hog1 . The involvement of MAPK in the biocontrol of Trichoderma has attracted much attention.…”

Section: Introductionmentioning

confidence: 99%

“…They belong to the socalled pathogenicity Fus3, cell-wall integrity Slt2, and osmoregulatory Hog1. 16 The involvement of MAPK in the biocontrol of Trichoderma has attracted much attention. Deletion of Fus3-MAPK gene tvk1 of T. virens enhanced biocontrol activity against R. solani and Pythium ultimum due to the production of more lytic enzymes.…”

Section: ■ Introductionmentioning

confidence: 99%

MAPK Cascades Mediating Biocontrol Activity of Trichoderma brevicrassum Strain TC967

Zhang

Zhuang

2022

J. Agric. Food Chem.

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Trichoderma brevicrassum strain TC967 is a novel biological control agent (BCA) against the plant pathogen Rhizoctonia solani and promotes plant growth. Mitogen-activated protein kinase (MAPK) cascades are involved in a variety of physiological functions of Trichoderma, but functions of each MAPK in regulating biocontrol have not been characterized in a single Trichoderma strain. In this study, we assembled and annotated the genome of strain TC967 and identified its three MAPK gene sequences. Functions of Fus3-, Slt2-, and Hog1-MAPK in strain TC967 were dissected. All three MAPKs were involved in hyphal growth. Hog1-MAPK was essential for conidiation and tolerance to saline/osmotic stress. Both Fus3- and Slt2-MAPK deletion mutants reduced cell-wall integrity (CWI) and increased the activities of chitinase and protease. The growth of Rhizoctoniasolani was further inhibited by volatile organic compounds (VOCs) and secondary metabolites produced by Fus3- and Slt2-MAPK deletion mutants, respectively. Biocontrol assays demonstrated that Fus3- and Slt2-MAPK deletion mutants were considerably more effective in disease control than the wild-type strain. RNA-seq analysis revealed that MAPK collectively played a major role in regulating biocontrol-related gene expressions, including the genes in charge of secondary metabolism, fungal cell wall-degrading enzymes (FCWDEs), and small secreted cysteine-rich proteins (SSCPs).

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Novel findings about the mode of action of the antifungal protein PeAfpA against Saccharomyces cerevisiae

Giner-Llorca,

Locascio,

del Real

et al. 2023

Appl Microbiol Biotechnol

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Antifungal proteins (AFPs) from filamentous fungi offer the potential to control fungal infections that threaten human health and food safety. AFPs exhibit broad antifungal spectra against harmful fungi, but limited knowledge of their killing mechanism hinders their potential applicability. PeAfpA from Penicillium expansum shows strong antifungal potency against plant and human fungal pathogens and stands above other AFPs for being active against the yeast Saccharomyces cerevisiae. We took advantage of this and used a model laboratory strain of S. cerevisiae to gain insight into the mode of action of PeAfpA by combining (i) transcriptional profiling, (ii) PeAfpA sensitivity analyses of deletion mutants available in the S. cerevisiae genomic deletion collection and (iii) cell biology studies using confocal microscopy. Results highlighted and confirmed the role of the yeast cell wall (CW) in the interaction with PeAfpA, which can be internalized through both energy-dependent and independent mechanisms. The combined results also suggest an active role of the CW integrity (CWI) pathway and the cAMP-PKA signalling in the PeAfpA killing mechanism. Besides, our studies revealed the involvement of phosphatidylinositol metabolism and the participation of ROX3, which codes for the subunit 19 of the RNA polymerase II mediator complex, in the yeast defence strategy. In conclusion, our study provides clues about both the killing mechanism of PeAfpA and the fungus defence strategies against the protein, suggesting also targets for the development of new antifungals. Key points • PeAfpA is a cell-penetrating protein with inhibitory activity against S. cerevisiae. • The CW integrity (CWI) pathway is a key player in the PeAfpA killing mechanism. • Phosphatidylinositol metabolism and ROX3 are involved in the yeast defence strategy.

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The GPI-Anchored GH76 Protein Dfg5 Affects Hyphal Morphology and Osmoregulation in the Mycoparasite Trichoderma atroviride and Is Interconnected With MAPK Signaling

Cited by 8 publications

References 70 publications

<i>Candida albicans</i> Mannosidases, Dfg5 and Dcw1, Are Required for Pathogenesis

<i>Candida albicans</i> Mannosidases, Dfg5 and Dcw1, Are Required for Pathogenesis

MAPK Cascades Mediating Biocontrol Activity of Trichoderma brevicrassum Strain TC967

Novel findings about the mode of action of the antifungal protein PeAfpA against Saccharomyces cerevisiae

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