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Introduction
This past decade, my work on the Nox (NADPH oxidase) complexes in the ascomycetes fungus Podospora anserina allowed me to study different stages of the fungal life cycle. Indeed, Nox control many processes in fungi such as fruiting body development, ascospore germination and appressorium development. As a geneticist and as a cell biologist, I am now investigating all these three processes through innovative approaches
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Publications
Publications (53)
In Podospora anserina as in many other Ascomycetes, ascospore germination is a regulated process that requires the breaking of dormancy. Despite its importance in survival and dispersal, ascospore germination in filamentous fungi has been poorly investigated, and little is known about its regulation and genetic control. We have designed a positive...
Sexual reproduction in Ascomycetes is well described in several model organisms such as Neurospora crassa or Podospora anserina. Deciphering the biological process of sexual reproduction (from the recognition between compatible partners to the formation of zygote) can be a major advantage to better control sexually reproducing pathogenic fungi. In...
In Podospora anserina as in many other ascomycetes , ascospore germination is a regulated process that requires breaking of dormancy. Despite its importance in survival and dispersal, ascospore germination in filamentous fungi has been poorly investigated and little is known about its regulation and genetic control. We have designed a positive gene...
Sexual reproduction is a key process influencing the evolution and adaptation of animals, plants, and many eukaryotic microorganisms, such as fungi. However, the sequential cell biology of fertilization and the associated nuclear dynamics after plasmogamy are poorly understood in filamentous fungi. Using histone-fluorescent parental isolates, we tr...
Sexual reproduction is a key process influencing the evolution and adaptation of animals, plants and many eukaryotic microorganisms, such as fungi. Mycologists have described the different fungal fruiting bodies, while geneticists have partly unravelled the regulation of sexual development. However, the sequential fungal cell biology of fertilisati...
Phytopathogenic and mycorrhizal fungi often penetrate living hosts by using appressoria and related structures. The differentiation of similar structures in saprotrophic fungi to penetrate dead plant biomass has seldom been investigated and has been reported only in the model fungus Podospora anserina. Here, we report on the ability of many saprotr...
Fungi are very successful microorganisms capable of colonizing virtually any ecological niche where they must constantly cope with competitors including fungi, bacteria and nematodes. We have shown previously that the ascomycete Podopora anserina exhibits Hyphal Interference (HI), an antagonistic response triggered by direct contact of competing fu...
Oxylipins are secondary messengers used universally in the living world for communication and defense. The paradigm is that they are produced enzymatically for the eicosanoids and non-enzymatically for the isoprostanoids. They are supposed to be degraded into volatile organic compounds (VOCs) and to participate in aroma production. Some such chemic...
The molecular pathways involved in the development of multicellular fruiting bodies in fungi are still not well known. Especially, the interplay between the mycelium, the female tissues and the zygotic tissues of the fruiting bodies is poorly documented. Here, we describe PM154, a new strain of the model ascomycetes Podospora anserina able to mate...
Background
Insects are well known vectors of human and animal pathogens and millions of people are killed by mosquito-borne diseases every year. The use of insecticides to target insect vectors has been hampered by the issues of toxicity to the environment and by the selection of resistant insects. Therefore, biocontrol strategies based on naturall...
The degradation of plant biomass is a major challenge towards the production of bio-based compounds and materials. As key lignocellulolytic enzyme producers, filamentous fungi represent a promising reservoir to tackle this challenge. Among them, the coprophilous ascomycete Podospora anserina has been used as a model organism to study various biolog...
NADPH oxidases (Nox) are membrane complexes that produce O2 (-) . Researches in mammals, plants and fungi highlight the involvement of Nox-generated ROS in cell proliferation, differentiation and defense. In mammals, the core enzyme gp91(phox) /Nox2 is associated with p22(phox) forming the flavocytochrome b558 ready for activation by a cytosolic co...
HMG-box domains used for Figure 2.
(TXT)
Code and accession numbers for proteins shown in Figure 2.
(DOC)
Alignment of HMG-box domains used for Figure 2.
(TXT)
Comparative organization around SpeSte11 orthologs in P. anserina and N. crassa. Orthologs were determined using FUNGIpath [52]. The gene names are enclosed in arrowed boxes indicating gene orientation. Orthologous genes are enclosed in boxes of identical color and connected with double arrows. Genes enclosed in white boxes do not have any ortholog...
Genomic Southern blots of HMG-box mutant strains probed with the hygromycin sequence. 1: DNA from ΔPahmg6 transformants # 1 and # 2 digested with KpnI. Both transformants displayed the expected pattern for homologous recombination. 2: DNA from ΔPahmg2 transformants # 1 and # 2 digested with KpnI. Only transformant # 2 displayed the expected pattern...
Electrophoretic mobility shift assays with PaHMG5 and mating-type target gene oligonucleotides. (A) Interaction of His tagged PaHMG5 (HMG5His) with probes corresponding to mat− mating-type target genes: Pa_1_8290 (MFM), Pa_1_9070 (PRE1) and Pa_1_7350 (7350). The probe is indicated below each panel with the sequence of its core HMG-box binding site....
Electrophoretic mobility shift assays with PaHMG5 and HMG-box gene oligonucleotides. Interaction of His tagged PaHMG5 (HMG5His) with probes corresponding to HMG-box genes: KEF1, mtHMG1, PaHMG2, PaHMG3, PaHMG4 and PaHMG6. Legend as in Figure S3 (A).
(TIFF)
Relative quantification of HMG-box gene and mating-type target gene transcription in ΔPahmg5 (ΔPa_1_13940) and WT strains.
(DOC)
Relative quantification of HMG-box gene and mating-type target gene transcription in Δkef1 (ΔPahmg9, ΔPa_1_7190) and WT strains.
(DOC)
Electrophoretic mobility shift reciprocal competition assays with PaHMG5 and HMG-box gene oligonucleotides. Legend as in Figure 8 (B). (A) Interaction of the HMG5His protein with the PaHMG2 probe and reciprocal competition between FMR1 and PaHMG2 oligonucleotides. HMG5His has a greater affinity for FMR1 than for PaHMG2 oligonucleotides, as indicate...
Color scheme used for Jalview.
(DOC)
Relative quantification of HMG-box gene and mating-type target gene transcription in ΔPahmg8 (ΔPa_6_4110) and WT strains.
(DOC)
Relative quantification of HMG-box gene and mating-type target gene transcription in Δmthmg1 (ΔPa_1_13340) and WT strains.
(DOC)
Phenotype of F. graminearum strains deleted for HMGB genes (adapted from [43]).
(DOC)
Relative quantification of HMG-box gene and mating-type target gene transcription in ΔPahmg6 (ΔPa_1_14230) and WT strains.
(DOC)
Oligonucleotide primers used for RT-qPCR.
(DOC)
Oligonucleotide primers used for HMG-box gene deletion.
(DOC)
Oligonucleotide primers used for HMG-box gene amplification.
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Complementation of P. anserina mutant strains.
(DOC)
Normalization genes (REF) used for RT-qPCR.
(DOC)
Oligonucleotides primers used for EMSA.
(DOC)
High-mobility group (HMG) B proteins are eukaryotic DNA-binding proteins characterized by the HMG-box functional motif. These transcription factors play a pivotal role in global genomic functions and in the control of genes involved in specific developmental or metabolic pathways. The filamentous ascomycete Podospora anserina contains 12 HMG-box ge...
Higher fungi, which comprise ascomycetes and basidiomycetes, play major roles in the biosphere. Their evolutionary success may be due to the extended dikaryotic stage of their life cycle, which is the basis for their scientific name: the Dikarya. Dikaryosis is maintained by similar structures, the clamp in basidiomycetes and the crozier in ascomyce...
Figure S1. Expression of homeobox genes during fruiting body maturation. Differential expression in space and time of the seven homeobox genes was assessed by microarray analysis in a time course of mycelium growth (Bidard and Silar, unpublished data) and perithecium maturation (Bidard and Berteaux-Lecellier, unpublished data). No difference in exp...
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...
Eumycetes fungi are a diverse group of organisms whose evolution is characterized by frequent changes in nutritional strategy
and the corresponding developmental programs. The reasons for this versatility are unknown. We previously discovered that
the NADPH oxidase Nox2 and the tetraspanin Pls1 are used in two radically different cell types to achi...
NADPH oxidases are enzymes that produce reactive oxygen species. Studies in mammals, plants and fungi have shown that they play important roles in differentiation, defence, host/pathogen interaction and mutualistic symbiosis. In this paper, we have identified a Podospora anserina mutant strain impaired for processes controlled by PaNox1 and PaNox2,...
Convergent evolution of trophic life style and morphological characters are very common in the fungal kingdom. Recently, we have shown that the same molecular machinery containing a tetraspanin and a NADPH oxidase has been recruited in two different fungal species for the same purpose (exiting from a melanized re-enforced cell at a focal weakened p...
In vitro, without Mediator, the association of general transcription factors (GTF) and RNA polymerase II (Pol II) in preinitiation complexes (PIC) occurs in an orderly fashion. In this work, we explore the in vivo function of Mediator in GTF recruitment to PIC. A direct interaction between Med11 Mediator head subunit and Rad3 TFIIH subunit was iden...
Correlation between Pretherapy Plasma HIV-1 RNA Levels as Determined by Amplicor HIV-1 Monitor Assay and SCA
Pretherapy samples from 157 participants were analyzed by both assays. 12 samples (triangles) had SCA values significantly lower than Amplicor and were excluded from further analysis. Remaining samples (n = 145, diamonds) were highly correla...
Schematic Describing Patient Inclusion for Study of HIV-1 Viremia in M98–863 Patients
(21 KB PDF)
We carried out gain-of-function mutagenesis screening and identified a mutant in which GAL4 induction led to both hyperplasia and apoptosis. The gene involved was identified as stonewall (stwl), a myb-related gene involved in germ cell proliferation and differentiation during oogenesis. As observed with dmyb, the ectopic expression of stwl(UY823) i...
Unlike mammalian Toll-like Receptors, the Drosophila Toll receptor does not interact directly with microbial determinants but is rather activated upon binding a cleaved form of the cytokine-like molecule Spatzle (Spz). During the immune response, Spz is thought to be processed by secreted serine proteases (SPs) present in the hemolymph that are act...
Septic injury triggers a rapid and widespread response in Drosophila adults that involves the up-regulation of many genes required to combat infection and for wound healing. Genome-wide expression profiling has already demonstrated that this response is controlled by signaling through the Toll, Imd, JAK-STAT and JNK pathways. Using oligonucleotide...
The Toll receptor was originally identified as an indispensable molecule for Drosophila embryonic development and subsequently as an essential component of innate immunity from insects to humans. Although in Drosophila the Easter protease processes the pro-Spätzle protein to generate the Toll ligand during development, the identification of the pro...
It is now established that genes involved in the execution of programmed cell death by apoptosis are relatively well conserved throughout evolution. However, the control of commitment to apoptosis exhibits some differences between organisms. In C. elegans, genetic studies have led to the identification of the ced genes (cell death) ced-3 and ced-4...
bcl-2 was the first regulator of apoptosis shown to be involved in oncogenesis. Subsequent studies in mammals, in the nematode and in Drosophila revealed wide evolutionary conservation of the regulation of apoptosis. Although dbok/debcl, a member of the bcl-2 gene family described in Drosophila, shows pro-apoptotic activities, no anti-apoptotic bcl...
Les genes impliques dans l'execution par apoptose de la mort cellulaire programmee ont ete conserves au cours de l'evolution. Toutefois, le controle de l'engagement dons ce processus varie selon les organismes. Chez Caenorhabditis elegans, l'approche genetique a permis de montrer que l'execution de l'opoptose est essentiellement assuree par le gene...
It is now established that the genes involved in the execution of programmed cell death by apoptosis are conserved throughout evolution. However, the control of commitment to apoptosis exhibits some differences between organisms. In C. elegans, genetic studies have led to the identification of the ced genes (cell death) ced-3 and ced-4 that are ess...
Studies of apoptosis in C. elegans have allowed the identification of three genes, ced-3, ced-4 and ced-9. Their products constitute the components of an induction pathway of apoptosis conserved in the nematode and mammals. In Drosophila, homologues have been found for CED-3, CED-4 and CED-9. CED-9 belongs to the Bcl-2 family which includes negativ...
Plasmid constructions and cloning All cloning were done using the Gateway Invitrogen cloning method. Wild-type MED11 gene was amplified from YPH499 genomic DNA using oligonucleotides matching the gene sequence initiation codon and following codons for the 5' forward primer and the stop codon and preceding codons for the 3' reverse primer. The oligo...
