Functions and regulation of the Nox family in the filamentous fungus <i>Podospora anserina</i>: a new role in cellulose degradation

Brun, Sylvain; Malagnac, Fabienne; Bidard, Frédérique; Lalucque, Hervé; Silar, Philippe

doi:10.1111/j.1365-2958.2009.06878.x

Cited by 110 publications

(148 citation statements)

References 53 publications

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“…Note that recovery of high numbers of mosaic perithecia would indicate a non-autonomous action of the mutated gene, i.e., only a fraction of the cells from the fruiting bodies being wild-type would be sufficient to promote normal development, possibly because the product of the wild-type gene would diffuse in the whole fruiting body. We have used this method to analyze mutants of the PaNox1, PaNoxR, PaMpk1, PaMpk2 and IDC1 genes [14,16,18,19]. The results confirmed and completed those of the grafting analysis: PaMpk1 and PaMpk2 are not required in the perithecium, but only in the mycelium; PaNox1 and PaNoxR are required only in the fruiting body and IDC1 is required in both the mycelium and the fruiting body.…”

Section: Fig (3) Grafting With Mutantssupporting

confidence: 66%

“…Conversely, perithecia will arrest at a precise stage if the gene is required at this stage in the zygotic tissues. We have used this method to analyze the PaMpk1, PaMpk2, PaNox1, PaNoxR, IDC1 mutants [14,18,19] confirming and completing previous data: PaMpk1, PaMpk2 are required only in the mycelium, PaNox1 and PaNoxR only in the maternal tissues of the fruiting bodies and IDC1 both in the mycelium and in the maternal tissues. We have also analyzed the pah5 beak defect [8] and showed that it was indeed due to a problem in the maternal tissues, as expected since the beak is made from maternal cells and not zygotic ones.…”

Section: Mosaic Analysis With the Mat Mutantmentioning

confidence: 51%

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Simple Genetic Tools to Study Fruiting Body Development in Fungi

Silar¹

2014

TOMYCJ

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Unlike for animal and plant development, the molecular processes involved in shaping the multicellular fruiting bodies of fungi are almost unknown. Especially, the interplay between the mycelium, the maternal tissues and zygotic ones are seldom investigated. Here, I summarized simple genetic methods that permit to allocate site(s) of action for genes whose mutations block fruiting body development. These involve the formation of genetic mosaics and grafting, as used for many decades to study embryo development in animals and plants. They are easily implemented without the requirement of complex equipment. Yet, they provide useful information when one wants to order genes into pathways acting during fruiting body production. Examples taken from the study of the model ascomycete Podospora anserina illustrate how they can be interpreted.

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Section: Fig (3) Grafting With Mutantssupporting

confidence: 66%

Section: Mosaic Analysis With the Mat Mutantmentioning

confidence: 51%

Simple Genetic Tools to Study Fruiting Body Development in Fungi

Silar¹

2014

TOMYCJ

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“…Briefly, strains carrying the investigated allele and either leu1-1 or lys2-1 were constructed and the ability of these strains to form a prototrophic mycelium was assayed on M2 minimal medium. Branching was observed as described (Arnaise et al 2001) and differentiation of appressorium-like structures was observed as reported (Brun et al 2009). Efficiency of cellulose degradation was measured as in Brun et al (2009), longevity as in Silar and Picard (1994), and fertility as in Rizet and Engelmann (1949).…”

Section: Phenotypic Analysismentioning

confidence: 99%

“…Branching was observed as described (Arnaise et al 2001) and differentiation of appressorium-like structures was observed as reported (Brun et al 2009). Efficiency of cellulose degradation was measured as in Brun et al (2009), longevity as in Silar and Picard (1994), and fertility as in Rizet and Engelmann (1949). Hyphal interference including peroxide production and cell death was assayed as described (Silar 2005) and DAB (diaminobenzidine) and NBT (nitroblue tetrazolium) staining to detect production of superoxides and peroxides, respectively, as in Malagnac et al (2004).…”

Section: Phenotypic Analysismentioning

confidence: 99%

A Non-Mendelian MAPK-Generated Hereditary Unit Controlled by a Second MAPK Pathway inPodospora anserina

et al. 2012

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The Podospora anserina PaMpk1 MAP kinase (MAPK) signaling pathway can generate a cytoplasmic and infectious element resembling prions. When present in the cells, this C element causes the crippled growth (CG) cell degeneration. CG results from the inappropriate autocatalytic activation of the PaMpk1 MAPK pathway during growth, whereas this cascade normally signals stationary phase. Little is known about the control of such prion-like hereditary units involved in regulatory inheritance. Here, we show that another MAPK pathway, PaMpk2, is crucial at every stage of the fungus life cycle, in particular those controlled by PaMpk1 during stationary phase, which includes the generation of C. Inactivation of the third P. anserina MAPK pathway, PaMpk3, has no effect on the development of the fungus. Mutants of MAPK, MAPK kinase, and MAPK kinase kinase of the PaMpk2 pathway are unable to present CG. This inability likely relies upon an incorrect activation of PaMpk1, although this MAPK is normally phosphorylated in the mutants. In PaMpk2 null mutants, hyphae are abnormal and PaMpk1 is mislocalized. Correspondingly, stationary phase differentiations controlled by PaMpk1 are defective in the mutants of the PaMpk2 cascade. Constitutive activation of the PaMpk2 pathway mimics in many ways its inactivation, including an effect on PaMpk1 localization. Analysis of double and triple mutants inactivated for two or all three MAPK genes undercover new growth and differentiation phenotypes, suggesting overlapping roles. Our data underscore the complex regulation of a prion-like element in a model organism.

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“…) NOXA and NOXB] are homologs of mammalian NOX2 and have been found in most ascomycetes. In contrast, NOX3, the homolog of mammalian NOX5, has been detected only in some fungi like Aspergillus terreus, Magnaporthe grisea, Podospora anserina, and several Fusarium species (Aguirre et al 2005;Scott and Eaton 2008;Brun et al 2009). Fungal NOX1 and NOX2 enzymes are regulated by the p67phox homolog NOR1 (NOX regulating, syn.…”

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confidence: 99%

New Insights Into the Roles of NADPH Oxidases in Sexual Development and Ascospore Germination in Sordaria macrospora

et al. 2014

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NADPH oxidase (NOX)-derived reactive oxygen species (ROS) act as signaling determinants that induce different cellular processes. To characterize NOX function during fungal development, we utilized the genetically tractable ascomycete Sordaria macrospora. Genome sequencing of a sterile mutant led us to identify the NADPH oxidase encoding nox1 as a gene required for fruiting body formation, regular hyphal growth, and hyphal fusion. These phenotypes are shared by Δnor1, lacking the NOX regulator NOR1. Further phenotypic analyses revealed a high correlation between increased ROS production and hyphal fusion deficiencies in Δnox1 and other sterile mutants. A genome-wide transcriptional profiling analysis of mycelia and isolated protoperithecia from wild type and Δnox1 revealed that nox1 inactivation affects the expression of genes related to cytoskeleton remodeling, hyphal fusion, metabolism, and mitochondrial respiration. Genetic analysis of Δnox2, lacking the NADPH oxidase 2 gene, Δnor1, and transcription factor deletion mutant Δste12, revealed a strict melanin-dependent ascospore germination defect, indicating a common genetic pathway for these three genes. We report that gsa3, encoding a G-protein a-subunit, and sac1, encoding cAMP-generating adenylate cyclase, act in a separate pathway during the germination process. The finding that cAMP inhibits ascospore germination in a melanin-dependent manner supports a model in which cAMP inhibits NOX2 activity, thus suggesting a link between both pathways. Our results expand the current knowledge on the role of NOX enzymes in fungal development and provide a frame to define upstream and downstream components of the NOX signaling pathways in fungi. D URING sexual reproduction, filamentous fungi generate complex fruiting bodies that contain and protect meiosporangia. We used the ascomycetous model fungus Sordaria macrospora to identify genes directly involved in fruiting body development (Kück et al. 2009;Engh et al. 2010;Kück et al. 2009). Due to its homothallic life style, S. macrospora is able to complete the sexual life cycle without the mating of strains with opposite sex, and therefore, fruiting body-deficient mutants can be recognized directly without the need for crossing experiments. In earlier work, we generated sterile mutants showing a developmental block after formation of young fruiting bodies (protoperithecia), but being unable to generate mature perithecia, and referred to these mutants as pro. Recently, we have applied next-generation genome re-sequencing to identify the genes affected in some of these mutants . Based on this approach, we now have characterized mutant pro32 and show that it carries a mutation in the nox1 gene encoding NAPDH oxidase 1 (NOX1).NADPH oxidase (NOX) enzymes are transmembrane proteins that are highly conserved among eukaryotes and produce reactive oxygen species (ROS) through the oxidation of NADPH (Lambeth 2004;Kawahara and Lambeth 2007). ROS have long been recognized as damaging agents due to uncontrolled oxidizing re...

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Functions and regulation of the Nox family in the filamentous fungus Podospora anserina: a new role in cellulose degradation

Cited by 110 publications

References 53 publications

Simple Genetic Tools to Study Fruiting Body Development in Fungi

Simple Genetic Tools to Study Fruiting Body Development in Fungi

A Non-Mendelian MAPK-Generated Hereditary Unit Controlled by a Second MAPK Pathway inPodospora anserina

New Insights Into the Roles of NADPH Oxidases in Sexual Development and Ascospore Germination in Sordaria macrospora

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