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the aspergillus nidulans peripheral er: disorganization by er stress and persistence during mitosis.the genetically amenable fungus aspergillus nidulans is well suited for cell biology studies involving the secretory pathway and its relationship with hyphal tip growth by apical extension. we exploited live-cell epifluorescence microscopy of the er labeled with the translocon component sec63, endogenously tagged with gfp, to study the organization of 'secretory' er domains. the sec63 a. nidulans er network includes brightly fluorescent peripheral strands and more faintly labeled nuclear envelop ...201323826221
xylh encodes a xylose/h+ symporter from the highly related yeast species debaryomyces fabryi and debaryomyces hansenii.the closely related yeasts debaryomyces fabryi and debaryomyces hansenii are excellent xylose consumers. we previously described the activity of a high-affinity xylose/h(+) symport from an industrial strain of d. hansenii subsequently reclassified as d. fabryi. we now report the identification of the gene encoding this permease, ay347871.2. this was retrieved from d. fabryi gdna using a degenerate primer pcr strategy, based on conserved regions from the amino acid sequences of three well-charact ...201323809840
functional characterisation of the non-essential protein kinases and phosphatases regulating aspergillus nidulans hydrolytic enzyme production.despite recent advances in the understanding of lignocellulolytic enzyme regulation, less is known about how different carbon sources are sensed and the signaling cascades that result in the adaptation of cellular metabolism and hydrolase secretion. therefore, the role played by non-essential protein kinases (npk) and phosphatases (npp) in the sensing of carbon and/or energetic status was investigated in the model filamentous fungus aspergillus nidulans.201323800192
a structural model of ppoa derived from saxs-analysis-implications for substrate conversion.in plants and mammals, oxylipins may be synthesized via multi step processes that consist of dioxygenation and isomerization of the intermediately formed hydroperoxy fatty acid. these processes are typically catalyzed by two distinct enzyme classes: dioxygenases and cytochrome p450 enzymes. in ascomycetes biosynthesis of oxylipins may proceed by a similar two-step pathway. an important difference, however, is that both enzymatic activities may be combined in a single bifunctional enzyme. these t ...201323797010
allopurinol and xanthine use different translocation mechanisms and trajectories in the fungal uapa transporter.in aspergillus nidulans uapa is a h(+)-driven transporter specific for xanthine, uric acid and several analogues. here, genetic and physiological evidence is provided showing that allopurinol is a high-affinity, low-capacity, substrate for uapa. surprisingly however, transport kinetic measurements showed that, uniquely among all recognized uapa substrates, allopurinol is transported by apparent facilitated diffusion and exhibits a paradoxical effect on the transport of physiological substrates. ...201323791789
metformin's performance in in vitro and in vivo genetic toxicology studies.metformin is a hypoglycemiant drug prescribed for the treatment and control of the type 2 diabetes mellitus. recently, the potential efficacy of this antidiabetic drug as an anticancer agent has been demonstrated in various mammalian cancer cells. this report evaluates the mutagenic as well as the recombinogenic potentials of the metformin drug in therapeutically relevant plasma concentrations (12.5 µm, 25.0 µm or 50.0 µm). since the loss of heterozygosity is a process associated with carcinogen ...201323788173
immobilization and biochemical properties of the enantioselective recombinant nstci esterase of aspergillus nidulans.the recombinant nstci a. nidulans esterase was adsorbed on accurel mp1000, where protein yield and immobilization efficiency were 42.48% and 81.94%, respectively. storage stability test at 4°c and rt showed 100% of residual activity after 40 days at both temperatures. the biocatalyst retains more than 70% of its initial activity after 3 cycles of repeated use. biochemical properties of this new biocatalyst were obtained. maximum activity was achieved at ph 11 and 30°c, while the best stability w ...201323781330
phospho-regulation and nucleocytoplasmic trafficking of crza in response to calcium and alkaline-ph stress in aspergillus nidulans.tolerance to abiotic stresses by microorganisms require of appropriate signalling and regulatory pathways. calcineurin phosphatases mediate calcium-dependent signalling pathways which are widely distributed among phylogeny. in saccharomyces cerevisiae, calcineurin mediates the post-translational modification of downstream effectors, most of them transcription factors, being the best-characterized calcineurin-regulated zinc-finger factor 1, crz1p. here we study the signalling process of crza, a f ...201323772954
analysis of a conserved cellulase transcriptional regulator reveals inducer-independent production of cellulolytic enzymes in neurospora crassa.cellulose is recalcitrant to deconstruction to glucose for use in fermentation strategies for biofuels and chemicals derived from lignocellulose. in neurospora crassa, the transcriptional regulator, clr-2, is required for cellulolytic gene expression and cellulose deconstruction. to assess conservation and divergence of cellulase gene regulation between fungi from different ecological niches, we compared clr-2 function with its ortholog (clrb) in the distantly related species, aspergillus nidula ...201323766336
the spt-ada-gcn5 acetyltransferase (saga) complex in aspergillus nidulans.a mutation screen in aspergillus nidulans uncovered mutations in the acdx gene that led to altered repression by acetate, but not by glucose. acdx of a. nidulans is highly conserved with spt8p of saccharomyces cerevisiae, and since spt8p is a component of the spt-ada-gcn5 acetyltransferase (saga) complex, the saga complex may have a role in acetate repression in a. nidulans. we used a bioinformatic approach to identify genes encoding most members of the saga complex in a. nidulans, and a proteom ...201323762321
co-immobilization of fungal endo-xylanase and α-l-arabinofuranosidase in glyoxyl agarose for improved hydrolysis of arabinoxylan.plant cell-wall arabinoxylans have a complex structure that requires the action of a pool of debranching (arabinofuranosidases) and depolymerizing enzymes (endo-xylanase). two aspergillus nidulans strains over-secreting endo-xylanase and arabinofuranosidase were inoculated in defined 2% maltose-minimum medium resulting in the simultaneously production of these enzymes. to study the synergistic hydrolysis was used arabinoxylan with 41% of arabinose and 59% of xylose residues. thus, it was adopted ...201323756760
uridylation prevents 3' trimming of oligoadenylated mrnas.degradation of mrnas is usually initiated by deadenylation, the shortening of long poly(a) tails to oligo(a) tails of 12-15 as. deadenylation leads to decapping and to subsequent 5' to 3' degradation by xrn proteins, or alternatively 3' to 5' degradation by the exosome. decapping can also be induced by uridylation as shown for the non-polyadenylated histone mrnas in humans and for several mrnas in schizosaccharomyces pombe and aspergillus nidulans. here we report a novel role for uridylation in ...201323748567
highly glucose tolerant β-glucosidase from aspergillus unguis: nii 08123 for enhanced hydrolysis of biomass.aspergillus unguis nii-08123, a filamentous fungus isolated from soil, was found to produce β-glucosidase (bgl) activity with high glucose tolerance. cultivation of the fungus in different carbon sources resulted in the secretion of different isoforms of the enzyme. a low molecular weight isoform, which retained ~60 % activity in the presence of 1.5 m glucose, was purified to homogeneity and the purified enzyme exhibited a temperature and ph optima of 60 °c and 6, respectively. the k(m) and v(ma ...201323732694
strain improvement by overexpression of the laea gene in monascus pilosus for the production of monascus-fermented rice.monascus species have been used to produce fermented rice called monascus-fermented rice (mfr). to improve a monascus strain via activation of secondary metabolite (sm) gene clusters for use in the production of mfr, we overexpressed an ortholog of the laea gene, which encodes a global positive regulator of secondary metabolism under the control of the strong heterologous aspergillus nidulans alca promoter in monascus pilosus. the oe::laea transformant produced more sms, including those not dete ...201323727802
live-cell imaging of aspergillus nidulans autophagy: rab1 dependence, golgi independence and er involvement.we exploited the amenability of the fungus aspergillus nidulans to genetics and live-cell microscopy to investigate autophagy. upon nitrogen starvation, gfp-atg8-containing pre-autophagosomal puncta give rise to cup-shaped phagophores and circular (0.9-μm diameter) autophagosomes that disappear in the vicinity of the vacuoles after their shape becomes irregular and their gfp-atg8 fluorescence decays. this 'autophagosome cycle' gives rise to characteristic cone-shaped traces in kymographs. autoph ...201323722157
acute inactivation of the aspergillus nidulans golgi membrane fusion machinery: correlation of apical extension arrest and tip swelling with cisternal disorganization.the mechanisms governing traffic across the golgi are incompletely understood. we studied, by live-cell microscopy, the consequences of disorganizing the aspergillus nidulans golgi, using an extended set of fluorescent protein markers to resolve early from late cisternae. the early golgi syntaxin sedv(sed) (5) and the rabo(rab) (1) regulatory gtpase play essential roles in secretion, cooperating in the er-golgi interface. following a temperature shift-up 'on-the-stage', hyphae carrying engineere ...201323714354
molecular genetic characterization of the biosynthesis cluster of a prenylated isoindolinone alkaloid aspernidine a in aspergillus nidulans.aspernidine a is a prenylated isoindolinone alkaloid isolated from the model fungus aspergillus nidulans. a genome-wide kinase knockout library of a. nidulans was examined, and it was found that a mitogen-activated protein kinase gene, mpka, deletion strain produces aspernidine a. targeted gene deletions were performed in the kinase deletion background to identify the gene cluster for aspernidine a biosynthesis. intermediates were isolated from mutant strains which provided information about the ...201323706169
neurospora crassa nkin2, a kinesin-3 motor, transports early endosomes and is required for polarized growth.biological motors are molecular nanomachines, which convert chemical energy into mechanical forces. the combination of mechanoenzymes with structural components, such as the cytoskeleton, enables eukaryotic cells to overcome entropy, generate molecular gradients, and establish polarity. hyphae of filamentous fungi are among the most polarized cells, and polarity defects are most obvious. here, we studied the role of the kinesin-3 motor, nkin2, in neurospora crassa. we found that nkin2 localizes ...201323687116
differences in composition of honey samples and their impact on the antimicrobial activities against drug multiresistant bacteria and pathogenic fungi.antibiotic multiresistant microbes represent a challenging problem. because honey has a potent antibacterial property, the antimicrobial effects of different honey samples against multiresistant pathogens and their compositions were investigated.201323684665
genetic bypass of aspergillus nidulans crza function in calcium homeostasis.after dephosphorylation by the phosphatase calcineurin, the fungal transcription factor crza enters the nucleus and activates the transcription of genes responsible for calcium homeostasis and many other calcium-regulated activities. a lack of crza confers calcium-sensitivity to the filamentous fungus aspergillus nidulans. to further understand calcium signaling in filamentous fungi and to identify genes that interact genetically with crza, we selected for mutations that were able to suppress cr ...201323665873
base composition and nucleosome density in exonic and intronic regions in genes of the filamentous ascomycetes aspergillus nidulans and aspergillus oryzae.we sequenced nucleosomal dna fragments of the filamentous ascomycetes aspergillus nidulans and aspergillus oryzae and then mapped those sequences on their genomes. we compared the gc content and nucleosome density in the exonic and intronic regions in the genes of a. nidulans and a. oryzae. although the gc content and nucleosome density in the exonic regions tended to be higher than those in the intronic regions, the difference in the distribution of the gc content was more notable than that of ...201323664982
identification of metabolic pathways influenced by the g-protein coupled receptors gprb and gprd in aspergillus nidulans.heterotrimeric g-protein-mediated signaling pathways play a pivotal role in transmembrane signaling in eukaryotes. our main aim was to identify signaling pathways regulated by a. nidulans gprb and gprd g-protein coupled receptors (gpcrs). when these two null mutant strains were compared to the wild-type strain, the δgprb mutant showed an increased protein kinase a (pka) activity while growing in glucose 1% and during starvation. in contrast, the δgprd has a much lower pka activity upon starvatio ...201323658706
conf and conj contribute to conidia germination and stress response in the filamentous fungus aspergillus nidulans.light induces various responses in fungi including formation of asexual and sexual reproductive structures. the formation of conidia in the filamentous fungus aspergillus nidulans is regulated by red and blue light receptors. expression of conidia associated con genes, which are widely spread in the fungal kingdom, increases upon exposure to light. we have characterized the light-inducible conf and conj genes of a. nidulans which are homologs of con-6 and con-10 of neurospora crassa. con genes a ...201323644150
β-glucan synthase gene overexpression and β-glucans overproduction in pleurotus ostreatus using promoter swapping.mushroom β-glucans are potent immunological stimulators in medicine, but their productivities are very low. in this study, we successfully improved its production by promoter engineering in pleurotus ostreatus. the promoter for β-1,3-glucan synthase gene (gls) was replaced by the promoter of glyceraldehyde-3-phosphate dehydrogenase gene of aspergillus nidulans. the homologous recombination fragment for swapping gls promoter comprised five segments, which were fused by two rounds of combined touc ...201323637884
pyrg is required for maintaining stable cellular uracil level and normal sporulation pattern under excess uracil stress in aspergillus nidulans.tight control of the intracellular uracil level is believed to be important to reduce the occurrence of uracil incorporation into dna. the pyrg gene of aspergillus nidulans encodes orotidine 5'-phosphate decarboxylase, which catalyzes the conversion of orotidine monophosphate (omp) to uridine monophosphate (ump). in this study, we found that pyrg is critical for maintaining uracil at a low concentration in a. nidulans cells in the presence of exogenous uracil. excess uracil and its derivatives h ...201323633078
an efficient system for heterologous expression of secondary metabolite genes in aspergillus nidulans.fungal secondary metabolites (sms) are an important source of medically valuable compounds. genome projects have revealed that fungi have many sm biosynthetic gene clusters that are not normally expressed. to access these potentially valuable, cryptic clusters, we have developed a heterologous expression system in aspergillus nidulans . we have developed an efficient system for amplifying genes from a target fungus, placing them under control of a regulatable promoter, transferring them into a. ...201323621425
comprehensive annotation of secondary metabolite biosynthetic genes and gene clusters of aspergillus nidulans, a. fumigatus, a. niger and a. oryzae.secondary metabolite production, a hallmark of filamentous fungi, is an expanding area of research for the aspergilli. these compounds are potent chemicals, ranging from deadly toxins to therapeutic antibiotics to potential anti-cancer drugs. the genome sequences for multiple aspergilli have been determined, and provide a wealth of predictive information about secondary metabolite production. sequence analysis and gene overexpression strategies have enabled the discovery of novel secondary metab ...201323617571
vertebral osteomyelitis and epidural abscess due to aspergillus nidulans resulting in spinal cord compression: case report and literature review.vertebral osteomyelitis caused by aspergillus nidulans is rare and usually affects immunocompromised patients. this report presents a case of thoracic vertebral osteomyelitis with epidural abscesses due to a. nidulans in a 40-year-old immunocompetent female who presented with back pain, numbness and weakness of both lower limbs. magnetic resonance imaging demonstrated osteomyelitis involving the thoracic (t)1-t3 vertebral bodies with epidural abscesses, resulting in spinal compression. the patie ...201323569017
water recycle as a must: decolorization of textile wastewaters by plant-associated fungi.textile dye effluents are among the most problematic pollutants because of their toxicity on several organisms and ecosystems. low cost and ecocompatible bioremediation processes offer a promising alternative to the conventional and aspecific physico-chemical procedures adopted so far. here, microorganisms resident on three real textile dyeing effluent were isolated, characterized, and tested for their decolorizing performances. although able to survive on these real textile-dyeing wastewaters, ...201423553673
recombinant aspergillus β-galactosidases as a robust glycomic and biotechnological tool.galactosidases are widespread enzymes that are used for manifold applications, including production of prebiotics, biosynthesis of different transgalactosylated products, improving lactose tolerance and in various analytical approaches. the nature of these applications often require galactosidases to be present in a purified form with clearly defined properties, including precisely determined substrate specificities, low sensitivity to inhibitors, and high efficiency and stability under distinct ...201424037406
in vitro and in silico analysis of the aspergillus nidulans dna-crea repressor interactions.the crea protein mediates carbon catabolite repression in the fungus aspergillus nidulans. its dna-binding domain belongs to the cys2-his2 class, binding specifically to a 5' syggrg 3' nucleotide sequence. emsa experiments showed that the crea(g27d) mutation resulted in a 30-fold increase of the kdiss, and footprinting revealed a altered pattern of protein/dna contacts. we modeled the crea and the crea(g27d) complexes in silico. a 15 ns molecular dynamics simulation of the solvated crea(g27d) an ...201424125468
the interplay between the gata transcription factors area, the global nitrogen regulator and areb in fusarium fujikuroi.nitrogen metabolite repression (nmr) in filamentous fungi is controlled by the gata transcription factors area and areb. while area mainly acts as a positive regulator of nmr-sensitive genes, the role of areb is not well understood. we report the characterization of areb and its interplay with area in the gibberellin-producing fungus fusarium fujikuroi. the areb locus produces three different transcripts that each code for functional proteins fully complementing the areb deletion mutant that inf ...201424286256
functional analysis of the c-ii subgroup killer toxin-like chitinases in the filamentous ascomycete aspergillus nidulans.chitinases are hydrolytic enzymes responsible for chitin polymer degradation. fungal chitinases belong exclusively to glycoside hydrolases family 18 and they are categorized into three phylogenetic groups (a, b and c), which are further divided into subgroups (a-ii to a-v, b-i to b-v and c-i to c-ii). subgroup c chitinases display similarity with the α/β-subunit of the zymocin yeast killer toxin produced by kluyveromyces lactis, suggesting a role of these enzymes in fungal-fungal interactions. i ...201424384382
insights into dynamic mitotic chromatin organization through the nima kinase suppressor sonc, a chromatin-associated protein involved in the dna damage response.the nuclear pore complex proteins sona and sonb, the orthologs of mammalian rae1 and nup98, respectively, were identified in aspergillus nidulans as cold-sensitive suppressors of a temperature-sensitive allele of the essential mitotic nima kinase (nima1). subsequent analyses found that sonb1 mutants exhibit temperature-dependent dna damage sensitivity. to understand this pathway further, we performed a genetic screen to isolate additional conditional dna damage-sensitive suppressors of nima1. we ...201424214344
establishing a versatile golden gate cloning system for genetic engineering in fungi.the corn pathogen ustilago maydis is a well-studied fungal model organism. along with a broad set of experimental tools, versatile strategies for the generation of gene replacement mutants by homologous recombination in u. maydis have been developed. nevertheless, the production of corresponding linear dna constructs still constitutes a time-limiting step. to overcome this bottleneck, various resistance cassette modules were adopted for use with the so-called golden gate cloning strategy. these ...201424211735
the aspergillus nidulans atm kinase regulates mitochondrial function, glucose uptake and the carbon starvation response.mitochondria supply cellular energy and also perform a role in the adaptation to metabolic stress. in mammals, the ataxia-telangiectasia mutated (atm) kinase acts as a redox sensor controlling mitochondrial function. subsequently, transcriptomic and genetic studies were utilized to elucidate the role played by a fungal atm homolog during carbon starvation. in aspergillus nidulans, atma was shown to control mitochondrial function and glucose uptake. carbon starvation responses that are regulated ...201424192833
the nima kinase is required to execute stage-specific mitotic functions after initiation of mitosis.the g2-m transition in aspergillus nidulans requires the nima kinase, the founding member of the nek kinase family. inactivation of nima results in a late g2 arrest, while overexpression of nima is sufficient to promote mitotic events independently of cell cycle phase. endogenously tagged nima-gfp has dynamic mitotic localizations appearing first at the spindle pole body and then at nuclear pore complexes before transitioning to within nuclei and the mitotic spindle and back at the spindle pole ...201424186954
weta is required for conidiogenesis and conidium maturation in the ascomycete fungus fusarium graminearum.fusarium graminearum, a prominent fungal pathogen that infects major cereal crops, primarily utilizes asexual spores to spread disease. to understand the molecular mechanisms underlying conidiogenesis in f. graminearum, we functionally characterized the f. graminearum ortholog of aspergillus nidulans weta, which has been shown to be involved in conidiogenesis and conidium maturation. deletion of f. graminearum weta did not alter mycelial growth, sexual development, or virulence, but the weta del ...201424186953
the cop9 signalosome is involved in the regulation of lipid metabolism and of transition metals uptake in saccharomyces cerevisiae.the cop9 signalosome (csn) is a highly conserved eukaryotic protein complex which regulates the cullin ring family of ubiquitin ligases and carries out a deneddylase activity that resides in subunit 5 (csn5). whereas csn activity is essential for the development of higher eukaryotes, several unicellular fungi including the budding yeast saccharomyces cerevisiae can survive without a functional csn. nevertheless, the budding yeast csn is biochemically active and deletion mutants of each of its su ...201424164706
the n-terminus region of the putative c2h2 transcription factor ada1 harbors a species-specific activation motif that regulates asexual reproduction in fusarium verticillioides.fusarium verticillioides is an important plant pathogenic fungus causing maize ear and stalk rots. in addition, the fungus is directly associated with fumonisin contamination of food and feeds. here, we report the functional characterization of ada1, a putative cys2-his2 zinc finger transcription factor with a high level of similarity to aspergillus nidulans flbc, which is required for the activation of the key regulator of conidiation brla. ada1 is predicted to encode a protein with two dna bin ...201424161731
characterization of aspergillus nidulans α-glucan synthesis: roles for two synthases and two amylases.cell walls are essential for fungal survival and growth. fungal walls are ∼ 90% carbohydrate, mostly types not found in humans, making them promising targets for anti-fungal drug development. echinocandins, which inhibit the essential β-glucan synthase, are already clinically available. in contrast, α-glucan, another abundant fungal cell wall component has attracted relatively little research attention because it is not essential for most fungi. aspergillus nidulans has two α-glucan synthases (a ...201424308872
astpt catalyses both reverse n1- and regular c2 prenylation of a methylated bisindolyl benzoquinone.prenylated bisindolyl benzoquinones exhibit interesting biological activities, such as antidiabetic or anti-hiv activities. a number of these compounds, including asterriquinones, have been isolated from aspergillus terreus. in this study, we identified two putative genes by genome mining, ateg_09980 and ateg_00702, which share high sequence similarity with the known bisindolyl benzoquinone prenyltransferase tdib from aspergillus nidulans. the coding sequences were cloned and overexpressed in e. ...201424302698
transport assays in filamentous fungi: kinetic characterization of the uapc purine transporter of aspergillus nidulans.transport assays allow the direct kinetic analysis of a specific transporter by measuring apparent km and vmax values, and permit the characterization of substrate specificity profiles through competition assays. in this protocol we describe a rapid and easy method for performing uptake assays in the model filamentous ascomycete aspergillus nidulans. our method makes use of a. nidulans germinating conidiospores at a defined morphological stage in which most transporters show maximal expression, ...201424355588
linkage of oxidative stress and mitochondrial dysfunctions to spontaneous culture degeneration in aspergillus nidulans.filamentous fungi including mushrooms frequently and spontaneously degenerate during subsequent culture maintenance on artificial media, which shows the loss or reduction abilities of asexual sporulation, sexuality, fruiting, and production of secondary metabolites, thus leading to economic losses during mass production. to better understand the underlying mechanisms of fungal degeneration, the model fungus aspergillus nidulans was employed in this study for comprehensive analyses. first, linkag ...201424345786
recent advances in genome mining of secondary metabolite biosynthetic gene clusters and the development of heterologous expression systems in aspergillus nidulans.fungi are prolific producers of secondary metabolites (sms) that show a variety of biological activities. recent advances in genome sequencing have shown that fungal genomes harbor far more sm gene clusters than are expressed under conventional laboratory conditions. activation of these "silent" gene clusters is a major challenge, and many approaches have been taken to attempt to activate them and, thus, unlock the vast treasure chest of fungal sms. this review will cover recent advances in geno ...201424342965
investigating aspergillus nidulans secretome during colonisation of cork cell walls.cork, the outer bark of quercus suber, shows a unique compositional structure, a set of remarkable properties, including high recalcitrance. cork colonisation by ascomycota remains largely overlooked. herein, aspergillus nidulans secretome on cork was analysed (2de). proteomic data were further complemented by microscopic (sem) and spectroscopic (atr-ftir) evaluation of the colonised substrate and by targeted analysis of lignin degradation compounds (uplc-hrms). data showed that the fungus forme ...201424316358
spread and change in stress resistance of shiga toxin-producing escherichia coli o157 on fungal colonies.to elucidate the effect of fungal hyphae on the behaviour of shiga toxin-producing escherichia coli (stec) o157, the spread and change in stress resistance of the bacterium were evaluated after coculture with 11 species of food-related fungi including fermentation starters. spread distances of stec o157 varied depending on the co-cultured fungal species, and the motile bacterial strain spread for longer distances than the non-motile strain. the population of stec o157 increased when co-cultured ...201423919289
inhibition of cereal rust fungi by both class i and ii defensins derived from the flowers of nicotiana alata.defensins are a large family of small, cysteine-rich, basic proteins, produced by most plants and plant tissues. they have a primary function in defence against fungal disease, although other functions have been described. this study reports the isolation and characterization of a class i secreted defensin (nad2) from the flowers of nicotiana alata, and compares its antifungal activity with the class ii defensin (nad1) from n. alata flowers, which is stored in the vacuole. nad2, like all other c ...201424015961
a novel thermophilic endo-β-1,4-mannanase from aspergillus nidulans xz3: functional roles of carbohydrate-binding module and thr/ser-rich linker region.the gene man5xz3 from aspergillus nidulans xz3 encodes a multimodular β-mannanase of glycoside hydrolase family 5 that consists of a family 1 carbohydrate-binding module (cbm1), a thr/ser-rich linker region, and a catalytic domain. recombinant man5xz3 and its two truncated derivatives, man5δcbm (removing the cbm1) and man5δcl (removing both the cbm1 and linker region), were produced in pichia pastoris and showed significant variance in the secondary structure. the three enzymes had similar bioch ...201423900799
molecular cloning, sequence analysis and expression of a ghf 43 xylanase from aspergillus niger in escherichia coli.a new xylanase gene (xyn43a) from aspergillus niger xz-3s was cloned and expressed in escherichia coli bl21-codonplus (de3)-ril. the coding region of the gene was separated by only one intron 86 bp in length. it encoded 318 amino acid residues of a protein with a calculated molecular weight (mw) of 33.47 kda plus a signal peptide of 19 amino acids. the amino acid sequence of the xyn43a gene showed 77.56% amino acid identity to a. nidulans xylanase, and the phylogenetic tree analysis revealed tha ...201425742974
development of rflp-pcr method for the identification of medically important aspergillus species using single restriction enzyme mwoi.in this study we attempted to modify the pcr-rflp method using restriction enzyme mwoi for the identification of medically important aspergillus species. our subjects included nine standard aspergillus species and 205 aspergillus isolates of approved hospital acquired infections and hospital indoor sources. first of all, aspergillus isolates were identified in the level of species by using morphologic method. a twenty four hours culture was performed for each isolates to harvest aspergillus myce ...201425242934
expression, purification, crystallization and preliminary x-ray diffraction analysis of aspergillus terreus endo-β-1,4-glucanase from glycoside hydrolase family 12.endoglucanases are important enzymes that are involved in the modification and degradation of cellulose. filamentous fungi such as aspergillus terreus are effective biomass degraders in nature owing to their capacity to produce an enzymatic arsenal of glycoside hydrolases, including endoglucanase from glycoside hydrolase family 12 (gh12). the a. terreus gh12 endoglucanase was cloned and overexpressed in a. nidulans, purified and crystallized. a single crystal was obtained from a solution consist ...201424637772
figa, a putative homolog of low-affinity calcium system member fig1 in saccharomyces cerevisiae, is involved in growth and asexual and sexual development in aspergillus nidulans.calcium-mediated signaling pathways are widely employed in eukaryotes and are implicated in the regulation of diverse biological processes. in saccharomyces cerevisiae, at least two different calcium uptake systems have been identified: the high-affinity calcium influx system (hacs) and the low-affinity calcium influx system (lacs). compared to the hacs, the lacs in fungi is not well known. in this study, figa, a homolog of the lacs member fig1 from s. cerevisiae, was functionally characterized ...201424376003
down-regulation of sidb gene by use of rna interference in aspergillus nidulans.introduction of the rna interference (rnai) machinery has guided the researchers to discover the function of essential vital or virulence factor genes in the microorganisms such as fungi. in the filamentous fungus aspergillus nidulans, the gene sidb plays an essential role in septation, conidiation and vegetative hyphal growth. in the present study, we benefited from the rnai strategy for down-regulating a vital gene, sidb, in the fungus a. nidulans.201424375164
a novel thermostable xylanase gh10 from malbranchea pulchella expressed in aspergillus nidulans with potential applications in biotechnology.the search for novel thermostable xylanases for industrial use has intensified in recent years, and thermophilic fungi are a promising source of useful enzymes. the present work reports the heterologous expression and biochemical characterization of a novel thermostable xylanase (gh10) from the thermophilic fungus malbranchea pulchella, the influence of glycosylation on its stability, and a potential application in sugarcane bagasse hydrolysis.201425788980
using aspergillus nidulans to identify antifungal drug resistance mutations.systemic fungal infections contribute to at least 10% of deaths in hospital settings. most antifungal drugs target ergosterol (polyenes) or its biosynthetic pathway (azoles and allylamines), or beta-glucan synthesis (echinocandins). antifungal drugs that target proteins are prone to the emergence of resistant strains. identification of genes whose mutations lead to targeted resistance can provide new information on those pathways. we used aspergillus nidulans as a model system to exploit its tra ...201424363365
in vitro photodynamic inactivation of plant-pathogenic fungi colletotrichum acutatum and colletotrichum gloeosporioides with novel phenothiazinium photosensitizers.the increasing tolerance to currently used fungicides in both clinical and agricultural areas is of great concern. the nonconventional light-based approach of antimicrobial photodynamic treatment (apdt) is a promising alternative to conventional fungicides. we evaluated the effects of apdt with four phenothiazinium derivatives (methylene blue [mb], new methylene blue n [nmbn], toluidine blue o [tbo], and the novel pentacyclic phenothiazinium photosensitizer [ps] s137) on conidia of three fungal ...201424362436
the transcription factor bcltf1 regulates virulence and light responses in the necrotrophic plant pathogen botrytis cinerea.botrytis cinerea is the causal agent of gray mold diseases in a range of dicotyledonous plant species. the fungus can reproduce asexually by forming macroconidia for dispersal and sclerotia for survival; the latter also participate in sexual reproduction by bearing the apothecia after fertilization by microconidia. light induces the differentiation of conidia and apothecia, while sclerotia are exclusively formed in the absence of light. the relevance of light for virulence of the fungus is not o ...201424415947
inhibition of tau aggregation by three aspergillus nidulans secondary metabolites: 2,ω-dihydroxyemodin, asperthecin, and asperbenzaldehyde.the aggregation of the microtubule-associated protein tau is a significant event in many neurodegenerative diseases including alzheimer's disease. the inhibition or reversal of tau aggregation is therefore a potential therapeutic strategy for these diseases. fungal natural products have proven to be a rich source of useful compounds having wide varieties of biological activity. we have screened aspergillus nidulans secondary metabolites containing aromatic ring structures for their ability to in ...201424414310
a microscopy-based screen employing multiplex genome sequencing identifies cargo-specific requirements for dynein velocity.the timely delivery of membranous organelles and macromolecules to specific locations within the majority of eukaryotic cells depends on microtubule-based transport. here we describe a screening method to identify mutations that have a critical effect on intracellular transport and its regulation using mutagenesis, multicolor-fluorescence microscopy, and multiplex genome sequencing. this screen exploits the filamentous fungus aspergillus nidulans, which has many of the advantages of yeast molecu ...201424403603
interactive exploration of integrated biological datasets using context-sensitive workflows.network inference utilizes experimental high-throughput data for the reconstruction of molecular interaction networks where new relationships between the network entities can be predicted. despite the increasing amount of experimental data, the parameters of each modeling technique cannot be optimized based on the experimental data alone, but needs to be qualitatively assessed if the components of the resulting network describe the experimental setting. candidate list prioritization and validati ...201424600467
application of a new dual localization-affinity purification tag reveals novel aspects of protein kinase biology in aspergillus nidulans.filamentous fungi occupy critical environmental niches and have numerous beneficial industrial applications but devastating effects as pathogens and agents of food spoilage. as regulators of essentially all biological processes protein kinases have been intensively studied but how they regulate the often unique biology of filamentous fungi is not completely understood. significant understanding of filamentous fungal biology has come from the study of the model organism aspergillus nidulans using ...201424599037
rational domain swaps reveal insights about chain length control by ketosynthase domains in fungal nonreducing polyketide synthases.a facile genetic methodology in the filamentous fungus aspergillus nidulans allowed exchange of the starter unit acp transacylase (sat) domain in the nonreduced polyketide synthase (nr-pks) afoe of the asperfuranone pathway with the sat domains from 10 other nr-pkss. the newly created hybrid with the nr-pks an3386 is able to accept a longer starter unit in place of the native substrate to create a novel aromatic polyketide in vivo.201424593241
velc positively controls sexual development in aspergillus nidulans.fungal development and secondary metabolism is intimately associated via activities of the fungi-specific velvet family proteins including vea, vosa, velb and velc. among these, velc has not been characterized in aspergillus nidulans. in this study, we characterize the role of velc in asexual and sexual development in a. nidulans. the velc mrna specifically accumulates during the early phase of sexual development. the deletion of velc leads to increased number of conidia and reduced production o ...201424587098
genetic characterization of the neurospora crassa molybdenum cofactor biosynthesis.molybdenum (mo) is a trace element that is essential for important cellular processes. to gain biological activity, mo must be complexed in the molybdenum cofactor (moco), a pterin derivative of low molecular weight. moco synthesis is a multi-step pathway that involves a variable number of genes in eukaryotes, which are assigned to four steps of eukaryotic moco biosynthesis. moco biosynthesis mutants lack any moco-dependent enzymatic activities, including assimilation of nitrate (plants and fung ...201424569084
multiple nuclear localization signals mediate nuclear localization of the gata transcription factor area.the aspergillus nidulans gata transcription factor area activates transcription of nitrogen metabolic genes in response to nitrogen limitation and is known to accumulate in the nucleus during nitrogen starvation. sequence analysis of area revealed multiple nuclear localization signals (nlss), five putative classical nlss conserved in fungal area orthologs but not in the saccharomyces cerevisiae functional orthologs gln3p and gat1p, and one putative noncanonical rrx33rxr bipartite nls within the ...201424562911
nsdd is a key repressor of asexual development in aspergillus nidulans.asexual development (conidiation) of the filamentous fungus aspergillus nidulans occurs via balanced activities of multiple positive and negative regulators. for instance, flug (+) and sfga (-) govern upstream regulation of the developmental switch, and brla (+) and vosa (-) control the progression and completion of conidiation. to identify negative regulators of conidiation downstream of flug-sfga, we carried out multicopy genetic screens using sfga deletion strains. after visually screening >1 ...201424532783
regulatory mutations affecting sulfur metabolism induce environmental stress response in aspergillus nidulans.mutations in the cysb, sconb and sconc genes affect sulfur metabolism in aspergillus nidulans in different ways. the cysb mutation blocks synthesis of cysteine by the main pathway and leads to a shortage of this amino acid. the sconb and sconc mutations affect subunits of the scf ubiquitin ligase complex, which inactivates the metr transcription factor in the presence of an excess of cysteine. in effect, both cysb and scon mutations lead to permanent derepression of metr-dependent genes. we comp ...201424513272
new branches in the degradation pathway of monochlorocatechols by aspergillus nidulans: a metabolomics analysis.a collective view of the degradation of monochlorocatechols in fungi is yet to be attained, though these compounds are recognised as key degradation intermediates of numerous chlorinated aromatic hydrocarbons, including monochlorophenols. in the present contribution we have analysed the degradation pathways of monochlorophenols in aspergillus nidulans using essentially metabolomics. degradation intermediates herein identified included those commonly reported (e.g. 3-chloro-cis,cis-muconate) but ...201424509097
furocoumarins and coumarins photoinactivate colletotrichum acutatum and aspergillus nidulans fungi under solar radiation.the increasing tolerance to currently-used fungicides is a major problem both in clinical and agricultural areas leading to an urgent need for the development of novel antifungal strategies. this study investigated the in vitro antimicrobial photo treatment (apt) of conidia of the plant-pathogenic fungus colletotrichum acutatum and the ascomycete aspergillus nidulans with the furocoumarins 8-methoxypsoralen (8-mop) and isopimpinellin, and a mixture of two coumarins (7-methoxy coumarin and citrop ...201424509069
enhanced cellulase producing mutants developed from heterokaryotic aspergillus strain.a heterokaryon 28, derived through protoplast fusion between aspergillus nidulans and aspergillus tubingensis (dal8), was subjected cyclic mutagenesis followed by selection on increasing levels of 2-deoxy glucose (2-dg) as selection marker. the derived deregulated cellulase hyper producing mutant '64', when compared to fusant 28, produced 9.83, 7.8, 3.2, 4.2 and 19.74 folds higher endoglucanase, β-glucosidase, cellobiohydrolase, fpase and xylanase, respectively, under shake cultures. the sequenc ...201424491293
control of reactive oxygen species (ros) production through histidine kinases in aspergillus nidulans under different growth conditions.sensor histidine kinases (hks) are important factors that control cellular growth in response to environmental conditions. the expression of 15 hks from aspergillus nidulans was analyzed by quantitative real-time pcr under vegetative, asexual, and sexual growth conditions. most hks were highly expressed during asexual growth. all hk gene-disrupted strains produced reactive oxygen species (ros). three hks are involved in the control of ros: hysa was the most abundant under the restricted oxygen c ...201424490133
cloning and characterization of a dynein light chain gene from puccinia striiformis f. sp. tritici.stripe rust is one of the most serious wheat diseases worldwide. the fungus puccinia striiformis f. sp. tritici (pst), the causal agent of this disease, is an obligate biotrophic basidiomycete fungus. numerous studies have shown that dyneins play important roles during fungal growth and propagation. however, knowledge is limited regarding the function of dyneins in pst. in this study, we cloned the dynein light chain gene psdlc1 from pst and characterized its expression. the function of psdlc1 w ...201424470306
mitotic regulation of fungal cell-to-cell connectivity through septal pores involves the nima kinase.intercellular bridges are a conserved feature of multicellular organisms. in multicellular fungi, cells are connected directly via intercellular bridges called septal pores. using aspergillus nidulans, we demonstrate for the first time that septal pores are regulated to be opened during interphase but closed during mitosis. septal pore-associated proteins display dynamic cell cycle-regulated locations at mature septa. of importance, the mitotic nima kinase locates to forming septa and surprising ...201424451264
liaison alcaline: pals entice non-endosomal escrts to the plasma membrane for ph signaling.the alkaline ph-responsive pal/rim signal transduction pathway mediating regulation of gene expression by ambient ph has been extensively studied in aspergillus nidulans and saccharomyces cerevisiae. in a. nidulans, palh, pali, palf, palc, pala and palb are required for the proteolytic activation of the executing transcription factor pacc. although necessary, pal proteins are insufficient to transmit the signal, which additionally requires escrt-i, ii and vps20 with snf7 in escrt-iii. although t ...201425460796
the velvet a orthologue vel1 of trichoderma reesei regulates fungal development and is essential for cellulase gene expression.trichoderma reesei is the industrial producer of cellulases and hemicellulases for biorefinery processes. their expression is obligatorily dependent on the function of the protein methyltransferase lae1. the aspergillus nidulans orthologue of lae1--laea--is part of the velvet protein complex consisting of laea, vea and velb that regulates secondary metabolism and sexual as well as asexual reproduction. here we have therefore investigated the function of vel1, the t. reesei orthologue of a. nidul ...201425386652
gene homozygosis and mitotic recombination induced by camptothecin and irinotecan in aspergillus nidulans diploid cells.mitotic recombination is a process involved in carcinogenesis which can lead to genetic loss through the loss of heterozygosity. the recombinogenic potentials of two anticancer drugs topoisomerase i inhibitors, camptothecin (cpt) and irinotecan (cpt-11), were evaluated in the present study. the homozygotization assay, which assess the induction of mitotic recombination and gene homozygosis, as well as the heterozygous a757//ut448 diploid strain of aspergillus nidulans were employed. the three no ...201425337669
neurospora crassa female development requires the pacc and other signal transduction pathways, transcription factors, chromatin remodeling, cell-to-cell fusion, and autophagy.using a screening protocol we have identified 68 genes that are required for female development in the filamentous fungus neurospora crassa. we find that we can divide these genes into five general groups: 1) genes encoding components of the pacc signal transduction pathway, 2) other signal transduction pathway genes, including genes from the three n. crassa map kinase pathways, 3) transcriptional factor genes, 4) autophagy genes, and 5) other miscellaneous genes. complementation and rip studies ...201425333968
mutations in proteins of the conserved oligomeric golgi complex affect polarity, cell wall structure, and glycosylation in the filamentous fungus aspergillus nidulans.we have described two aspergillus nidulans gene mutations, designated podb1 (polarity defective) and swop1 (swollen cell), which cause temperature-sensitive defects during polarization. mutant strains also displayed unevenness and abnormal thickness of cell walls. un-polarized or poorly-polarized mutant cells were capable of establishing normal polarity after a shift to a permissive temperature, and mutant hyphae shifted from permissive to restrictive temperature show wall and polarity abnormali ...201425312861
the aspergillus nidulans signalling mucin msba regulates starvation responses, adhesion and affects cellulase secretion in response to environmental cues.in the heterogeneous semi-solid environment naturally occupied by lignocellulolytic fungi the majority of nutrients are locked away as insoluble plant biomass. hence, lignocellulolytic fungi must actively search for, and attach to, a desirable source of nutrients. during growth on lignocellulose a period of carbon deprivation provokes carbon catabolite derepression and scavenging hydrolase secretion. subsequently, starvation and/or contact sensing was hypothesized to play a role in lignocellulos ...201425294314
protein phosphatase 2a (pp2a) regulatory subunits para and paba orchestrate septation and conidiation and are essential for pp2a activity in aspergillus nidulans.protein phosphatase 2a (pp2a) is a major intracellular protein phosphatase that regulates multiple aspects of cell growth and metabolism. different activities of pp2a and subcellular localization are determined by its regulatory subunits. here we identified and characterized the functions of two protein phosphatase regulatory subunit homologs, para and paba, in aspergillus nidulans. our results demonstrate that para localizes to the septum site and that deletion of para causes hyperseptation, wh ...201425280816
fgflbd regulates hyphal differentiation required for sexual and asexual reproduction in the ascomycete fungus fusarium graminearum.fusarium graminearum is a filamentous fungal plant pathogen that infects major cereal crops. the fungus produces both sexual and asexual spores in order to endure unfavorable environmental conditions and increase their numbers and distribution across plants. in a model filamentous fungus, aspergillus nidulans, early induction of conidiogenesis is orchestrated by the fluffy genes. the objectives of this study were to characterize fluffy gene homologs involved in conidiogenesis and their mechanism ...201425277408
transcriptional regulation of fksa, a β-1,3-glucan synthase gene, by the apses protein stua during aspergillus nidulans development.the temporal and spatial regulation of β-1,3-glucan synthesis plays an important role in morphogenesis during fungal growth and development. northern blot analysis showed that the transcription of fksa, the gene encoding β-1,3-glucan synthase in aspergillus nidulans, was cell-cycle-dependent and increased steadily over the duration of the vegetative period, but its overall expression during the asexual and sexual stages was fairly constant up until the time of transcription cessation. in an a. n ...201425359270
characterization of the mutagenic spectrum of 4-nitroquinoline 1-oxide (4-nqo) in aspergillus nidulans by whole genome sequencing.4-nitroquinoline 1-oxide (4-nqo) is a highly carcinogenic chemical that induces mutations in bacteria, fungi, and animals through the formation of bulky purine adducts. 4-nqo has been used as a mutagen for genetic screens and in both the study of dna damage and dna repair. in the model eukaryote aspergillus nidulans, 4-nqo-based genetic screens have been used to study diverse processes, including gene regulation, mitosis, metabolism, organelle transport, and septation. early work during the 1970 ...201425352541
penicillium antifungal protein (paf) is involved in the apoptotic and autophagic processes of the producer penicillium chrysogenum.paf, which is produced by the filamentous fungus pencicillium chrysogenum, is a small antifungal protein, triggering ros-mediated apoptotic cell death in aspergillus nidulans. in this work, we provide information on the function of paf in the host p. chrysogenum considering that carbon-starving cultures of the δpaf mutant strain showed significantly reduced apoptosis rates in comparison to the wild-type (wt) strain. moreover, the addition of paf to the δpaf strain resulted in a twofold increase ...201425261948
the vip1 inositol polyphosphate kinase family regulates polarized growth and modulates the microtubule cytoskeleton in fungi.microtubules (mts) are pivotal for numerous eukaryotic processes ranging from cellular morphogenesis, chromosome segregation to intracellular transport. execution of these tasks requires intricate regulation of mt dynamics. here, we identify a new regulator of the schizosaccharomyces pombe mt cytoskeleton: asp1, a member of the highly conserved vip1 inositol polyphosphate kinase family. inositol pyrophosphates generated by asp1 modulate mt dynamic parameters independent of the central +tip eb1 a ...201425254656
flow cytometry of microencapsulated colonies for genetics analysis of filamentous fungi.the analysis of filamentous fungi by flow cytometry has been impossible to date due to their filamentous nature and size. in this work, we have developed a method that combines single-spore microencapsulation and large-particle flow cytometry as a powerful alternative for the genetic analysis of filamentous fungi. individual spores were embedded in monodisperse alginate microparticles and incubated in the appropriate conditions. growth could be monitored by light or fluorescent microscopy and co ...201425239104
microtubule plus end-tracking proteins play critical roles in directional growth of hyphae by regulating the dynamics of cytoplasmic microtubules in aspergillus nidulans.cytoplasmic microtubules (mts) serve as a rate-limiting factor for hyphal tip growth in the filamentous fungus aspergillus nidulans. we hypothesized that this function depended on the mt plus end-tracking proteins (+tips) including the eb1 family protein eba that decorated the mt plus ends undergoing polymerization. the ebaδ mutation reduced colony growth and the mutant hyphae appeared in an undulating pattern instead of exhibiting unidirectional growth in the control. these phenotypes were enha ...201425213466
the role of +tips in directional tip expansion.aspergillus nidulans is an ideal model to study nuclear migration and intracellular transport by dynein and kinesin owing to its long neuron-like hyphae, conserved transport mechanisms, and powerful genetics. in this organism, as in other filamentous fungi, microtubules have been implicated in patterning cell shape through polarized tip growth - the hallmark mode of growth that generates the elongated hyphae. exactly how microtubules regulate tip growth is incompletely understood and remains a f ...201425213368
an isoprenylation and palmitoylation motif promotes intraluminal vesicle delivery of proteins in cells from distant species.the c-terminal ends of small gtpases contain hypervariable sequences which may be posttranslationally modified by defined lipid moieties. the diverse structural motifs generated direct proteins towards specific cellular membranes or organelles. however, knowledge on the factors that determine these selective associations is limited. here we show, using advanced microscopy, that the isoprenylation and palmitoylation motif of human rhob (-cincckvl) targets chimeric proteins to intraluminal vesicle ...201425207810
expression of aspergillus nidulans phy gene in nicotiana benthamiana produces active phytase with broad specificities.a full-length phytase gene (phy) of aspergillus nidulans was amplified from the cdna library by polymerase chain reaction (pcr), and it was introduced into a bacterial expression vector, pet-28a. the recombinant protein (rphy-e, 56 kda) was overexpressed in the insoluble fraction of escherichia coli culture, purified by ni-nta resin under denaturing conditions and injected into rats as an immunogen. to express a. nidulans phytase in a plant, the full-length of phy was cloned into a plant express ...201425192284
hydroxamate production as a high affinity iron acquisition mechanism in paracoccidioides spp.iron is a micronutrient required by almost all living organisms, including fungi. although this metal is abundant, its bioavailability is low either in aerobic environments or within mammalian hosts. as a consequence, pathogenic microorganisms evolved high affinity iron acquisition mechanisms which include the production and uptake of siderophores. here we investigated the utilization of these molecules by species of the paracoccidioides genus, the causative agents of a systemic mycosis. it was ...201425157575
a dually located multi-hmg-box protein of aspergillus nidulans has a crucial role in conidial and ascospore germination.seven hmg-box proteins of aspergillus nidulans have been identified in the genomic databases. three of these have the characteristics of non-specific dna-binding proteins. one of these, an1267 (hmbb), comprises one canonical hmg-box in its c-terminus and upstream of the canonical box two structurally related boxes, to be called shadow-hmg-boxes. this protein defines, together with the podospora anserina mthmg1, a clade of proteins present in the pezizomycotina, with orthologues in some of the ta ...201425156107
protein kinase c overexpression suppresses calcineurin-associated defects in aspergillus nidulans and is involved in mitochondrial function.in filamentous fungi, intracellular signaling pathways which are mediated by changing calcium levels and/or by activated protein kinase c (pkc), control fungal adaptation to external stimuli. a rise in intracellular ca2+ levels activates calcineurin subunit a (cnaa), which regulates cellular calcium homeostasis among other processes. pkc is primarily involved in maintaining cell wall integrity (cwi) in response to different environmental stresses. cross-talk between the ca2+ and pkc-mediated pat ...201425153325
metabolism of d-galactose is dispensable for the induction of the beta-galactosidase (bgad) and lactose permease (lacpa) genes in aspergillus nidulans.in this study, we analyze the expression of the aspergillus nidulans bgad-lacpa gene couple (encoding an intracellular beta-galactosidase and a lactose permease) in the presence of d-galactose. this monosaccharide can be catabolized via alternative, independent pathways in this model organism. the inductive capabilities of intermediates of the two alternative routes of d-galactose utilization were addressed in loss-of-function mutants defective in a defined step in one of the two pathways. in a ...201425145606
putative pmra and pmca are important for normal growth, morphogenesis and cell wall integrity, but not for viability in aspergillus nidulans.p-type ca(2+)-transporting atpases are ca(2+) pumps, extruding cytosolic ca(2+) to the extracellular environment or the intracellular ca(2+) store lumens. in budding yeast, pmr1 (plasma membrane atpase related), and pmc1 (plasma membrane calcium-atpase) cannot be deleted simultaneously for it to survive in standard medium. here, we deleted two putative ca(2+) pumps, designated anpmra and anpmca, from aspergillus nidulans, and obtained the mutants δanpmra and δanpmca, respectively. then, using δa ...201425118249
the phosphoproteome of aspergillus nidulans reveals functional association with cellular processes involved in morphology and secretion.we describe the first phosphoproteome of the model filamentous fungus aspergillus nidulans. phosphopeptides were enriched using titanium dioxide, separated using a convenient ultra-long reverse phase gradient, and identified using a "high-high" strategy (high mass accuracy on the parent and fragment ions) with higher-energy collisional dissociation. using this approach 1801 phosphosites, from 1637 unique phosphopeptides, were identified. functional classification revealed phosphoproteins were ov ...201425116090
restraint of the g2/m transition by the sr/rrm family mrna shuttling binding protein snxahrb1 in aspergillus nidulans.control of the eukaryotic g2/m transition by cdc2/cyclinb is tightly regulated by protein-protein interactions, protein phosphorylations, and nuclear localization of cdc2/cyclinb. we previously reported a screen, in aspergillus nidulans, for extragenic suppressors of nimx2(cdc2) that resulted in the identification of the cold-sensitive snxa1 mutation. we demonstrate here that snxa1 suppresses defects in regulators of the cdk1 mitotic induction pathway, including nimx2(cdc) (2), nime6(cyclinb), a ...201425104516
genome organization and assessment of high copy number and increased expression of pectinolytic genes from penicillium griseoroseum: a potential heterologous system for protein production.the fungus penicillium griseoroseum has the potential for application on an industrial scale as a host for the production of homologous and heterologous proteins, mainly because it does not produce some mycotoxins or secrete proteases under the growth conditions for pectinase production. however, for the fungus to be used effectively as an expression heterologous system, an understanding of the organization of its genome, as well as the mechanisms of gene expression and protein production, is re ...201425099254
high-yield recombinant xylanase production by aspergillus nidulans under pyridoxine limitation.the present study investigated the limitation of pyridoxine on an aspergillus nidulans culture that produces xylanase b (xynb) as a client enzyme and was unable to synthesize pyridoxine. this technique was used to limit cell growth and divert substrate to product formation for a surface grown culture that could be used in trickle bed reactors. it was observed that growth was limited when pyridoxine was absent, while enzyme production was unaffected. enzyme production was 1,026 u after 480 h of c ...201425085742
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