Modulation of calcineurin activity in <i>Aspergillus nidulans</i>: the roles of high magnesium concentrations and of transcriptional factor CrzA

Manoli, Maria‐Tsampika; Espeso, Eduardo A.

doi:10.1111/mmi.14221

Cited by 11 publications

(12 citation statements)

References 48 publications

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“…The procedures for nLC-MS/MS were previously described in Manoli and Espeso (2019). A 20-ml aliquot of extracellular protein precipitate was loaded onto a 10% polyacrylamide gel and allowed to run 1 cm into the resolving gel.…”

Section: Nlc-ms/ms Proteomicsmentioning

confidence: 99%

Proteomic Studies to Understand the Mechanisms of Peach Tissue Degradation by Monilinia laxa

et al. 2020

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Monilinia laxa is a necrotrophic plant pathogen able to infect and produce substantial losses on stone fruit. Three different isolates of M. laxa were characterized according to their aggressiveness on nectarines. M. laxa 8L isolate was the most aggressive on fruit, 33L isolate displayed intermediated virulence level, and 5L was classified as a weak aggressive isolate. Nectarine colonization process by the weak isolate 5L was strongly delayed. nLC-MS/MS proteomic studies using in vitro peach cultures provided data on exoproteomes of the three isolates at equivalent stages of brown rot colonization; 3 days for 8L and 33L, and 7 days for 5L. A total of 181 proteins were identified from 8L exoproteome and 289 proteins from 33L at 3 dpi, and 206 proteins were identified in 5L exoproteome at 7 dpi. Although an elevated number of proteins lacked a predicted function, the vast majority of proteins belong to OG group "metabolism", composed of categories such as "carbohydrate transport and metabolism" in 5L, and "energy production and conversion" most represented in 8L and 33L. Among identified proteins, 157 that carried a signal peptide were further examined and classified. Carbohydrate-active enzymes and peptidases were the main groups revealing different protein alternatives with the same function among isolates. Our data suggested a subset of secreted proteins as possible markers of differential virulence in more aggressive isolates, MlPG1 MlPME3, NEP-like, or endoglucanase proteins. A core-exoproteome among isolates independently of their virulence but time-dependent was also described. This core included several well-known virulence factors involved in host-tissue factors like cutinase, pectin lyases, and acid proteases. The secretion patterns supported the assumption that M. laxa deploys an extensive repertoire of proteins to facilitate the host infection and colonization and provided information for further characterization of M. laxa pathogenesis.

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Section: Nlc-ms/ms Proteomicsmentioning

confidence: 99%

Proteomic Studies to Understand the Mechanisms of Peach Tissue Degradation by Monilinia laxa

et al. 2020

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“…While the expression levels of three genes rcnA , chsB , and pmcA were decreased in the Δ crzA mutant than those in control, pmrA mRNA level was not changed in the Δ crzA mutant. These results indicate that CrzA supports rcnA and pmcA expression and this activity is conserved in other Aspergillus species [34,43]. Overall, these finding indicate that the roles of CrzA in cell wall biogenesis and stress response are conserved in Aspergillus species, but target genes of CrzA are diverse in Aspergillus species.…”

Section: Resultsmentioning

confidence: 61%

“…Crz1 is a C 2 H 2 -type transcription factor that is a key target for calcineurin in yeast and filamentous fungi [26]. Recent studies analyzed the function and domains of CrzA in two Aspergillus species A. fumigatus and A. nidulans [42,43]. CrzA from both species contains putative a calcineurin-binding domain (CBD), DNA-binding domain (DBD, C 2 H 2 domain), nuclear localization signal (NLS), and serine-rich region.…”

Section: Resultsmentioning

confidence: 99%

“…However, A. flavus CrzA does not contain an NES at the N-terminus, unlike the A. fumigatus CrzA. The A. flavus CrzA protein also contains CBD and serine-rich regions that are essential for regulating the activity of CrzA [42,43]. The DBD, CBD, and NLS elements of the CrzA homologs in most Aspergillus spp are highly conserved, but the NES element is diverse in 19 Aspergillus species (Figure 1).…”

Section: Resultsmentioning

confidence: 99%

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Function of crzA in Fungal Development and Aflatoxin Production in Aspergillus flavus

Lim

Son

Lee

et al. 2019

Toxins

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The calcineurin pathway is an important signaling cascade for growth, sexual development, stress response, and pathogenicity in fungi. In this study, we investigated the function of CrzA, a key transcription factor of the calcineurin pathway, in an aflatoxin-producing fungus Aspergillus flavus (A. flavus). To examine the role of the crzA gene, crzA deletion mutant strains in A. flavus were constructed and their phenotypes, including fungal growth, spore formation, and sclerotial formation, were examined. Absence of crzA results in decreased colony growth, the number of conidia, and sclerocia production. The crzA-deficient mutant strains were more susceptible to osmotic pressure and cell wall stress than control or complemented strains. Moreover, deletion of crzA results in a reduction in aflatoxin production. Taken together, these results demonstrate that CrzA is important for differentiation and mycotoxin production in A. flavus.

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“…Candidate proteins were identified using available P. rubens protein databases at NCBI (National Center for Biotechnology Information). Sample preparation for nLC-MS was as previously described [ 41 ]. The mass spectrometry data from three or two biological replicates were analyzed searching against P. rubens Wisconsin strain 54-1255 [ 42 ] database (12,771 protein sequences), using the Sequest search engine through Proteome Discoverer (version 1.4.1.14, Thermo Scientific) using default parameters.…”

Section: Methodsmentioning

confidence: 99%

Exploring the Extracellular Macromolecular Composition of Crude Extracts of Penicillium rubens Strain 212 for Elucidation Its Mode of Action as a Biocontrol Agent

Carreras¹,

Espeso

Gutiérrez-Docio

et al. 2020

JoF

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Penicillium rubens strain 212 (PO212) acts as an inducer of systemic resistance in tomato plants. The effect of crude extracellular extracts of PO212 on the soil-borne pathogen Fusarium oxysporum f. sp. lycopersici has been evaluated. Evidence of the involvement of soluble, thermo-labile, and proteinase-inactivated macromolecules present in PO212 crude extracts in the control of Fusarium vascular disease in tomato plants was found. Proteomic techniques and the availability of the access to the PO212 genome database have allowed the identification of glycosyl hydrolases, oxidases, and peptidases in these extracellular extracts. Furthermore, a bioassay-guided fractionation of PO212 crude extracellular extracts using an integrated membrane/solid phase extraction process was set up. This method enabled the separation of a PO212 crude extracellular extract of seven days of growth into four fractions of different molecular sizes and polarities: high molecular mass protein fraction >5 kDa, middle molecular mass protein fraction 5–1 kDa, low molecular mass metabolite fraction, and nutrients from culture medium (mainly glucose and minerals). The high and middle molecular mass protein fractions retained disease control activity in a way similar to that of the control extracts. Proteomic techniques have allowed the identification of nine putatively secreted proteins in the high molecular mass protein fraction matching those identified in the total crude extracts. Therefore, these enzymes are considered to be potentially responsible of the crude extracellular extract-induced resistance in tomato plants against F. oxysporum f. sp. lycopersici. Further studies are required to establish which of the identified proteins participate in the PO212’s action mode as a biocontrol agent.

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Modulation of calcineurin activity in Aspergillus nidulans: the roles of high magnesium concentrations and of transcriptional factor CrzA

Cited by 11 publications

References 48 publications

Proteomic Studies to Understand the Mechanisms of Peach Tissue Degradation by Monilinia laxa

Proteomic Studies to Understand the Mechanisms of Peach Tissue Degradation by Monilinia laxa

Function of crzA in Fungal Development and Aflatoxin Production in Aspergillus flavus

Exploring the Extracellular Macromolecular Composition of Crude Extracts of Penicillium rubens Strain 212 for Elucidation Its Mode of Action as a Biocontrol Agent

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