219 research outputs found

    Aspergillus nidulans pyrE and pyrF

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    Sequencing of a gene cloned by complementation of a pyr mutant in Glasgow strain G190 shows this mutant to be pyrE, encoding dihydroorotate dehydrogenase. This discovery necessitates a revision of the genetic map derived from crosses with this strain

    Distinct Roles for Intra- and Extracellular Siderophores during Aspergillus fumigatus Infection

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    Siderophore biosynthesis by the highly lethal mould Aspergillus fumigatus is essential for virulence, but non-existent in humans, presenting a rare opportunity to strategize therapeutically against this pathogen. We have previously demonstrated that A. fumigatus excretes fusarinine C and triacetylfusarinine C to capture extracellular iron, and uses ferricrocin for hyphal iron storage. Here, we delineate pathways of intra- and extracellular siderophore biosynthesis and show that A. fumigatus synthesizes a developmentally regulated fourth siderophore, termed hydroxyferricrocin, employed for conidial iron storage. By inactivation of the nonribosomal peptide synthetase SidC, we demonstrate that the intracellular siderophores are required for germ tube formation, asexual sporulation, resistance to oxidative stress, catalase A activity, and virulence. Restoration of the conidial hydroxyferricrocin content partially rescues the virulence of the apathogenic siderophore null mutant ΔsidA, demonstrating an important role for the conidial siderophore during initiation of infection. Abrogation of extracellular siderophore biosynthesis following inactivation of the acyl transferase SidF or the nonribosomal peptide synthetase SidD leads to complete dependence upon reductive iron assimilation for growth under iron-limiting conditions, partial sensitivity to oxidative stress, and significantly reduced virulence, despite normal germ tube formation. Our findings reveal distinct cellular and disease-related roles for intra- and extracellular siderophores during mammalian Aspergillus infection

    Spatio-temporal variability in underwater light climate in a turbid river-floodplain system. Driving factors and estimation using Secchi disc

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    The underwater light climate has important effects on primary producers. The aim of this research was to evaluate its variability in a turbid river-floodplain system. Photosynthetically active radiation (PAR) was measured in the Middle Paraná River during different hydrological phases to (a) analyse the photosynthetically active radiation attenuation coefficient (k) and euphotic depth (Zeu) as well as their associations with optically active components and (b) develop and evaluate indices and regression models based on Secchi disc (SD) measurements to estimate k and Zeu. Values of k were higher in the fluvial system than in the floodplain and during low-water stage than high-water stage. Particulate components controlled the light climate variability. Chromophoric dissolved organic matter and chlorophyll-a had significant effects during floods. The estimation of k and Zeu was sensitive to temporal but not to spatial variations. The highest prediction accuracy was observed when using specific non-linear regressions for each hydrological phase, especially for Zeu estimation (low stage: k = 1.76 × SD−0.80, Zeu = 2.62 × 1/SD−0.80; high stage: k = 2.04 × SD−0.53, Zeu = 2.26 × 1/SD−0.53). The indices k × SD and Zeu/SD were significantly different from those proposed for clear water environments. It is concluded that temporal variations should be considered when estimating k and Zeu in turbid river-floodplain systems because of the temporal heterogeneity in optically active components. Considering that ecological implication of the light climate depends on Zeu:depth ratio, we propose to estimate Zeu instead of k. Finally, indices proposed for clear water environments are not recommended to be applied to turbid environments.Fil: Mayora, Gisela Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; ArgentinaFil: Devercelli, Melina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; Argentin

    Community profiling and gene expression of fungal assimilatory nitrate reductases in agricultural soil

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    Although fungi contribute significantly to the microbial biomass in terrestrial ecosystems, little is known about their contribution to biogeochemical nitrogen cycles. Agricultural soils usually contain comparably high amounts of inorganic nitrogen, mainly in the form of nitrate. Many studies focused on bacterial and archaeal turnover of nitrate by nitrification, denitrification and assimilation, whereas the fungal role remained largely neglected. To enable research on the fungal contribution to the biogeochemical nitrogen cycle tools for monitoring the presence and expression of fungal assimilatory nitrate reductase genes were developed. To the ∼100 currently available fungal full-length gene sequences, another 109 partial sequences were added by amplification from individual culture isolates, representing all major orders occurring in agricultural soils. The extended database led to the discovery of new horizontal gene transfer events within the fungal kingdom. The newly developed PCR primers were used to study gene pools and gene expression of fungal nitrate reductases in agricultural soils. The availability of the extended database allowed affiliation of many sequences to known species, genera or families. Energy supply by a carbon source seems to be the major regulator of nitrate reductase gene expression for fungi in agricultural soils, which is in good agreement with the high energy demand of complete reduction of nitrate to ammonium

    Characterization of an Nmr Homolog That Modulates GATA Factor-Mediated Nitrogen Metabolite Repression in Cryptococcus neoformans

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    Nitrogen source utilization plays a critical role in fungal development, secondary metabolite production and pathogenesis. In both the Ascomycota and Basidiomycota, GATA transcription factors globally activate the expression of catabolic enzyme-encoding genes required to degrade complex nitrogenous compounds. However, in the presence of preferred nitrogen sources such as ammonium, GATA factor activity is inhibited in some species through interaction with co-repressor Nmr proteins. This regulatory phenomenon, nitrogen metabolite repression, enables preferential utilization of readily assimilated nitrogen sources. In the basidiomycete pathogen Cryptococcus neoformans, the GATA factor Gat1/Are1 has been co-opted into regulating multiple key virulence traits in addition to nitrogen catabolism. Here, we further characterize Gat1/Are1 function and investigate the regulatory role of the predicted Nmr homolog Tar1. While GAT1/ARE1 expression is induced during nitrogen limitation, TAR1 transcription is unaffected by nitrogen availability. Deletion of TAR1 leads to inappropriate derepression of non-preferred nitrogen catabolic pathways in the simultaneous presence of favoured sources. In addition to exhibiting its evolutionary conserved role of inhibiting GATA factor activity under repressing conditions, Tar1 also positively regulates GAT1/ARE1 transcription under non-repressing conditions. The molecular mechanism by which Tar1 modulates nitrogen metabolite repression, however, remains open to speculation. Interaction between Tar1 and Gat1/Are1 was undetectable in a yeast two-hybrid assay, consistent with Tar1 and Gat1/Are1 each lacking the conserved C-terminus regions present in ascomycete Nmr proteins and GATA factors that are known to interact with each other. Importantly, both Tar1 and Gat1/Are1 are suppressors of C. neoformans virulence, reiterating and highlighting the paradigm of nitrogen regulation of pathogenesis

    Cosying up to MoCo

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