Extracellular calcium triggers unique transcriptional programs and modulates staurosporine-induced cell death in Neurospora crassa

Gonçalves, A. Pedro; Monteiro, João; Lucchi, Chiara; Kowbel, David; Cordeiro, JM; Correia-de-Sá, Paulo; Rigden, Daniel J.; Glass, N. Louise; Videira, Arnaldo

doi:10.15698/mic2014.09.165

Cited by 8 publications

(6 citation statements)

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“…Ca 2+ is also involved in the increasing threshold of N. crassa to antifungals such as staurosporine. 44 Calcium plays a role in the mechanisms of plasma membrane remodeling in S. cerevisiae budding 45 and during cell fusion in N. crassa . 14,17 In this work, we report NCU10610 (Ca 2+ regulator with fig 1 domain) significantly repressed by chitosan.…”

Section: Resultsmentioning

confidence: 99%

Neurospora crassa transcriptomics reveals oxidative stress and plasma membrane homeostasis biology genes as key targets in response to chitosan

et al. 2016

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Chitosan is a natural polymer with antimicrobial activity. Chitosan causes plasma membrane permeabilization and induction of intracellular reactive oxygen species (ROS) in Neurospora crassa. We have determined the transcriptional profile of N. crassa to chitosan and identified the main gene targets involved in the cellular response to this compound. Global network analyses showed membrane, transport and oxidoreductase activity as key nodes affected by chitosan. Activation of oxidative metabolism indicates the importance of ROS and cell energy together with plasma membrane homeostasis in N. crassa response to chitosan. Deletion strain analysis of chitosan susceptibility pointed, NCU03639 encoding a class 3 lipase, involved in plasma membrane repair by lipid replacement and NCU04537 a MFS monosaccharide transporter related with assimilation of simple sugars, as main gene targets of chitosan. NCU10521, a glutathione S-transferase-4 involved in the generation of reducing power for scavenging intracellular ROS is also a determinant chitosan gene target. Ca2+ increased tolerance to chitosan in N. crassa. Growth of NCU10610 (fig 1 domain) and SYT1 (a synaptotagmin) deletion strains was significantly increased by Ca2+ in presence of chitosan. Both genes play a determinant role in N. crassa membrane homeostasis. Our results are of paramount importance for developing chitosan as antifungal.

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Section: Resultsmentioning

confidence: 99%

Neurospora crassa transcriptomics reveals oxidative stress and plasma membrane homeostasis biology genes as key targets in response to chitosan

et al. 2016

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“…Phospholipase C seems to be a pivotal player coordinating recruitment of Ca 2+ to the cytosol during staurosporine-induced cell death as cells lacking the phospolipase C gene plc-2 exhibit increased survival and a staurosporine-induced cytosolic Ca 2+ signature is abolished (Goncalves et al, 2014a ). The importance of extracellular Ca 2+ during staurosporine-induced fungal PCD is supported by the observation that cell death in N. crassa is exacerbated in Ca 2+ -free medium and inhibited when excess Ca 2+ is present (Gonçalves et al, 2014 ). Consistent with these observations, a similar protection from fungal PCD was observed by the presence of an excessive amount of extracellular Ca 2+ in occidiofungin-treated S. cerevisiae cells and chitosan-treated N. crassa cells (Lopez-Moya et al, 2016 ; Robinson et al, 2017 ).…”

Section: Molecular Mediators Of Regulated Cell Deathmentioning

confidence: 96%

Regulated Forms of Cell Death in Fungi

et al. 2017

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Cell death occurs in all domains of life. While some cells die in an uncontrolled way due to exposure to external cues, other cells die in a regulated manner as part of a genetically encoded developmental program. Like other eukaryotic species, fungi undergo programmed cell death (PCD) in response to various triggers. For example, exposure to external stress conditions can activate PCD pathways in fungi. Calcium redistribution between the extracellular space, the cytoplasm and intracellular storage organelles appears to be pivotal for this kind of cell death. PCD is also part of the fungal life cycle, in which it occurs during sexual and asexual reproduction, aging, and as part of development associated with infection in phytopathogenic fungi. Additionally, a fungal non-self-recognition mechanism termed heterokaryon incompatibility (HI) also involves PCD. Some of the molecular players mediating PCD during HI show remarkable similarities to major constituents involved in innate immunity in metazoans and plants. In this review we discuss recent research on fungal PCD mechanisms in comparison to more characterized mechanisms in metazoans. We highlight the role of PCD in fungi in response to exogenic compounds, fungal development and non-self-recognition processes and discuss identified intracellular signaling pathways and molecules that regulate fungal PCD.

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“…These observations prompted us to examine the transcriptional profile of N. crassa germlings grown in the absence of calcium in the external medium (http://www.ncbi.nlm.nih.gov/geo/; series record: GSE53013) (24). We narrowed our search to genes that showed decreased expression levels in the absence of calcium and encoded proteins containing transmembrane domains; 50 genes were identified that met these criteria (see Table S1 in the supplemental material).…”

Section: Identification Of New Genes Involved In Germling Fusionmentioning

confidence: 99%

“…Our strategy relied on the identification of genes encoding predicted transmembrane proteins and which showed a significant decrease in expression level in the absence of extracellular calcium (24). We screened 29 strains carrying deletions of candidate genes for defects in membrane merger and cell lysis during germling fusion.…”

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Identification and Characterization of LFD-2, a Predicted Fringe Protein Required for Membrane Integrity during Cell Fusion in Neurospora crassa

et al. 2015

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The molecular mechanisms of membrane merger during somatic cell fusion in eukaryotic species are poorly understood. In the filamentous fungus Neurospora crassa, somatic cell fusion occurs between genetically identical germinated asexual spores (germlings) and between hyphae to form the interconnected network characteristic of a filamentous fungal colony. In N. crassa, two proteins have been identified to function at the step of membrane fusion during somatic cell fusion: PRM1 and LFD-1. The absence of either one of these two proteins results in an increase of germling pairs arrested during cell fusion with tightly appressed plasma membranes and an increase in the frequency of cell lysis of adhered germlings. The level of cell lysis in ⌬Prm1 or ⌬lfd-1 germlings is dependent on the extracellular calcium concentration. An available transcriptional profile data set was used to identify genes encoding predicted transmembrane proteins that showed reduced expression levels in germlings cultured in the absence of extracellular calcium. From these analyses, we identified a mutant (lfd-2, for late fusion defect-2) that showed a calcium-dependent cell lysis phenotype. lfd-2 encodes a protein with a Fringe domain and showed endoplasmic reticulum and Golgi membrane localization. The deletion of an additional gene predicted to encode a low-affinity calcium transporter, fig1, also resulted in a strain that showed a calcium-dependent cell lysis phenotype. Genetic analyses showed that LFD-2 and FIG1 likely function in separate pathways to regulate aspects of membrane merger and repair during cell fusion. Membrane fusion plays an important role in intracellular trafficking of vesicles, exo-and endocytosis, fertilization, the entry of enveloped animal viruses into their host cell, and syncytium formation during developmental processes (1-3). In intracellular vesicle trafficking, SNARE proteins facilitate membrane merger by forming coiled-coil interactions between v-SNARES on vesicles and t-SNARES on target membranes to bring membranes into close proximity (1, 4). The mechanisms that direct cell-cell fusion are less well understood, although genetic screens have identified candidate molecules, termed fusogens, which are membrane-incorporated proteins that facilitate membrane fusion. For example, in Caenorhabditis elegans, two fusogenic glycoproteins, EFF-1 and AFF-1, are necessary and sufficient to fuse cells (5, 6), but their conservation is restricted to nematodes and related organisms (7).In the yeast Saccharomyces cerevisiae, a number of proteins that are involved in mating cell fusion have been identified (8)(9)(10)(11)(12)(13)(14). Mating pairs between strains bearing deletions of Prm1p or Fig1p show tightly appressed plasma membranes, indicating a defect in membrane fusion (10, 12). Mating pairs of prm1⌬ or fig1⌬ strains also show increased cell lysis, the frequency of which is influenced by the concentration of extracellular calcium (12, 15). However, Prm1p and Fig1p are not fusogens, as neither is necessary or sufficient f...

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Extracellular calcium triggers unique transcriptional programs and modulates staurosporine-induced cell death in Neurospora crassa

Cited by 8 publications

References 61 publications

Neurospora crassa transcriptomics reveals oxidative stress and plasma membrane homeostasis biology genes as key targets in response to chitosan

Neurospora crassa transcriptomics reveals oxidative stress and plasma membrane homeostasis biology genes as key targets in response to chitosan

Regulated Forms of Cell Death in Fungi

Identification and Characterization of LFD-2, a Predicted Fringe Protein Required for Membrane Integrity during Cell Fusion in Neurospora crassa

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