TitleAbstractYear(sorted descending)
biosynthesis of silver, gold and bimetallic nanoparticles using the filamentous fungus neurospora crassa.the development of production processes that can reduce the environmental impact, offer waste reduction and increase energy efficiency is an important step in the field of application of nanotechnology. in this work the filamentous fungus neurospora crassa was screened and found to be successful for the production of mono and bimetallic au/ag nanoparticles (nps). analysis by scanning electron microscopy (sem), energy dispersive x-ray spectroscopy (eds), and transmission electron microscopy (tem) ...201121087843
activation and localization of protein kinase c in neurospora crassa.the neurosporacrassa protein kinase c (npkc) is reported to be a regulator of light responsive genes. it phosphorylates the light receptor wc-1 and regulates the levels of the circadian clock protein frq and transcription of the light-induced albino-2 gene. in mammals, the conventional and novel isoforms of pkc are activated by diacylglycerol (dag), which induces pkc translocation from the cytoplasm to membranes. to investigate the interaction of npkc and dag in neurospora, we constructed a stra ...201121070858
population genomics and local adaptation in wild isolates of a model microbial eukaryote.elucidating the connection between genotype, phenotype, and adaptation in wild populations is fundamental to the study of evolutionary biology, yet it remains an elusive goal, particularly for microscopic taxa, which comprise the majority of life. even for microbes that can be reliably found in the wild, defining the boundaries of their populations and discovering ecologically relevant phenotypes has proved extremely difficult. here, we have circumvented these issues in the microbial eukaryote n ...201121282627
traffic of chitin synthase 1 (chs-1) to the spitzenkörper and developing septa in hyphae of neurospora crassa: actin dependence and evidence of distinct microvesicle populations.we describe the subcellular location of chitin synthase 1 (chs-1), one of seven chitin synthases in neurospora crassa. laser scanning confocal microscopy of growing hyphae showed chs-1-green fluorescent protein (gfp) localized conspicuously in regions of active wall synthesis, namely, the core of the spitzenkörper (spk), the apical cell surface, and developing septa. it was also present in numerous fine particles throughout the cytoplasm plus some large vacuoles in distal hyphal regions. althoug ...201121296914
carrefour mme. gras: a wild-isolated neurospora crassa strain that suppresses meiotic silencing by unpaired dna and uncovers a novel ascospore stability defect.ordinarily, rip-induced erg-3 mutant neurospora crassa ascospores and their erg(+) siblings do not differ in stability during long-term storage. consequently, the frequency of rip-induced erg-3 mutants remains about constant regardless of the time that has elapsed between ascospore harvest and germination. we found, however, that rip-induced erg-3 mutants were apparently selectively lost with time from among the ascospores stored from a cross with the wild-isolated carrefour mme. gras strain fro ...201121295150
circadian timekeeping in neurospora crassa and synechococcus first, the saprophytic eukaryote neurospora crassa and the photosynthetic prokaryote synechococcus elongatus may seem to have little in common. however, in both organisms a circadian clock organizes cellular biochemistry, and each organism lends itself to classical and molecular genetic investigations that have revealed a detailed picture of the molecular basis of circadian rhythmicity. in the present chapter, an overview of the molecular clockwork in each organism will be described, highligh ...201121819383
cofermentation of cellobiose and galactose by an engineered saccharomyces cerevisiae strain.we demonstrate improved ethanol yield and productivity through cofermentation of cellobiose and galactose by an engineered saccharomyces cerevisiae strain expressing genes coding for cellodextrin transporter (cdt-1) and intracellular +¦-glucosidase (gh1-1) from neurospora crassa. simultaneous fermentation of cellobiose and galactose can be applied to producing biofuels from hydrolysates of marine plant biomass.201121705527
expression and characterization of the neurospora crassa endoglucanase gh5-1.filamentous fungi secrete a wide range of enzymes, including cellulases and hemicellulases, with potential applications in the production of lignocellulosic biofuels. of the cellulolytic fungi, hypocrea jecorina (anamorph trichoderma reesei) is the best characterized in terms of cellulose degradation, but other cellulolytic fungi, such as the model filamentous fungus neurospora crassa, can serve a crucial role in building our knowledge about the fungal response to biomass due to the many molecul ...201120826217
involvement of a putative response regulator fgrrg-1 in osmotic stress response, fungicide resistance and virulence in fusarium graminearum.response regulator (rr) proteins are core elements of the high-osmolarity glycerol (hog) pathway, which plays an important role in the adaptation of fungi to a variety of environmental stresses. in this study, we constructed deletion mutants of two putative rr genes, fgrrg-1 and fgrrg-2, which are orthologues of neurospora crassa rrg-1 and rrg-2, respectively. the fgrrg-1 deletion mutant (δfgrrg1-6) showed increased sensitivity to osmotic stress mediated by nacl, kcl, sorbitol or glucose, and to ...201121535349
high-throughput production of gene replacement mutants in neurospora crassa.the model filamentous fungus neurospora crassa has been the focus of functional genomics studies for the past several years. a high-throughput gene knockout procedure has been developed and used to generate mutants for more than two-thirds of the ∼10,000 annotated n. crassa genes. yeast recombinational cloning was incorporated as an efficient procedure to produce all knockout cassettes. n. crassa strains with the δmus-51 or δmus-52 deletion mutations were used as transformation recipients in ord ...201121590421
Structural elements of the mitochondrial preprotein-conducting channel Tom40 dissolved by bioinformatics and mass spectrometry.Most mitochondrial proteins are imported into mitochondria from the cytosolic compartment. Proteins destined for the outer or inner membrane, the inter-membrane space, or the matrix are recognized and translocated by the TOM machinery containing the specialized protein import channel Tom40. The latter is a protein with ß-barrel shape, which is suggested to have evolved from a porin-type protein. To obtain structural insights in the absence of a crystal structure the membrane topology of Tom40 fr ...201121888892
self/nonself recognition in tuber melanosporum is not mediated by a heterokaryon incompatibility system.vegetative incompatibility is a widespread phenomenon in filamentous ascomycetes, which limits formation of viable heterokaryons. whether this phenomenon plays a role in maintaining the homokaryotic state of the hyphae during the vegetative growth of tuber spp. gene expression, polymorphism analysis as well as targeted in vitro experiments allowed us to test whether a heterokaryon incompatibility (hi) system operates in tuber melanosporum. hi is controlled by different genetic systems, often inv ...201122289772
the kinetics of changes in the fatty acid composition of neurospora crassa lipids after a temperature increase.the fatty acid composition of the total lipids, phospholipids and neutral lipids of log-phase shaker cultures of the bd (band) strain of neurospora crassa, were measured every 2 h for an 8-h period following a temperature increase from 22 to 40 degrees c. for purposes of comparison, the fatty acid composition was also measured when cultures were grown from inoculation at temperatures between 22 and 40 degrees c. in the phospholipids, the temperature jump produced, over a 4-6 hour span, a linear ...20113159434
The Neurospora crassa mutant Nc?Egt-1 identifies an ergothioneine biosynthetic gene and demonstrates that ergothioneine enhances conidial survival and protects against peroxide toxicity during conidial germination.Ergothioneine (EGT) is a histidine derivative with sulfur on the imidazole ring and a trimethylated amine; it is postulated to have an antioxidant function. Although EGT apparently is only produced by fungi and some prokaryotes, it is acquired by animals and plants from the environment, and is concentrated in animal tissues in cells with an EGT transporter. Monobromobimane derivatives of EGT allowed conclusive identification of EGT by LC/MS and the quantification of EGT in Colletotrichum gramini ...201122209968
the azoarcus group i intron ribozyme misfolds and is accelerated for refolding by atp-dependent rna chaperone proteins.structured rnas traverse complex energy landscapes that include valleys representing misfolded intermediates. in neurospora crassa and saccharomyces cerevisiae, efficient splicing of mitochondrial group i and ii introns requires the dead box proteins cyt-19 and mss116p, respectively, which promote folding transitions and function as general rna chaperones. to test the generality of rna misfolding and the activities of dead box proteins in vitro, here we measure native folding of a small group i ...201121878649
meiotic silencing in the homothallic fungus gibberella zeae.the homothallic ascomycete fungus gibberella zeae is an important pathogen on major cereal crops. the objective of this study was to determine whether meiotic silencing occurs in g. zeae. cytological studies demonstrated that gfp and rfp-fusion proteins were not detected during meiosis, both in heterozygous outcrosses and homozygous selfings. the deletion of rsp-1, a homologue used for studies on meiotic silencing of neurospora crassa, triggered abnormal ascospores from selfing, but outcrosses b ...201122115448
biochemical characterisation of a nadph-dependent carbonyl reductase from neurospora crassa reducing α- and β-keto esters.a gene encoding an nadph-dependent carbonyl reductase from neurospora crassa (nccr) was cloned and heterologously expressed in escherichia coli. the enzyme (nccr) was purified and biochemically characterised. nccr exhibited a restricted substrate spectrum towards various ketones, and the highest activity (468u/mg) was observed with dihydroxyacetone. however, nccr proved to be very selective in the reduction of different α- and β-keto esters. several compounds were converted to the corresponding ...201122113019
Identification of the CRE-1 cellulolytic regulon in Neurospora crassa.In filamentous ascomycete fungi, the utilization of alternate carbon sources is influenced by the zinc finger transcription factor CreA/CRE-1, which encodes a carbon catabolite repressor protein homologous to Mig1 from Saccharomyces cerevisiae. In Neurospora crassa, deletion of cre-1 results in increased secretion of amylase and ß-galactosidase.201121980519
Meiotic silencing by unpaired DNA is expressed more strongly in the early than the late perithecia of crosses involving most wild-isolated Neurospora crassa strains and in self-crosses of N. tetrasperma.Meiotic silencing by unpaired DNA is a presumed RNAi-mediated elimination of the transcripts of any gene that is not properly paired with a homolog in meiosis. Eighty wild-isolated strains of Neurospora crassa were classified into three types based on the apparent strength of meiotic silencing of the bml (ß-tubulin) and mei-3 genes in crosses with the ::Bml(r) and ::mei-3 tester strains. "OR" and "Sad" type wild-isolates, respectively, did or did not silence both the genes, whereas the "Esm" typ ...201122056520
evolution of multicopper oxidase genes in coprophilous and non-coprophilous members of the order sordariales.multicopper oxidases (mco) catalyze the biological oxidation of various aromatic substrates and have been identified in plants, insects, bacteria, and wood rotting fungi. in nature, they are involved in biodegradation of biopolymers such as lignin and humic compounds, but have also been tested for various industrial applications. in fungi, mcos have been shown to play important roles during their life cycles, such as in fruiting body formation, pigment formation and pathogenicity. coprophilous f ...201121966247
analysis of al-2 mutations in neurospora.the orange pigmentation of the fungus neurospora crassa is due to the accumulation of the xanthophyll neurosporaxanthin and precursor carotenoids. two key reactions in the synthesis of these pigments, the formation of phytoene from geranylgeranyl pyrophosphate and the introduction of +¦ cycles in desaturated carotenoid products, are catalyzed by two domains of a bifunctional protein, encoded by the gene al-2. we have determined the sequence of nine al-2 mutant alleles and analyzed the carotenoid ...201121818281
Catalytically active filaments - pyruvate decarboxylase from Neurospora crassa. pH-controlled oligomer structure and catalytic function.Pyruvate decarboxylase is a key enzyme in organisms whose energy metabolism is based on alcoholic fermentation. The enzyme catalyses the nonoxidative decarboxylation of 2-oxo acids in the presence of the cofactors thiamine diphosphate and magnesium ions. Pyruvate decarboxylase species from yeasts and plant seeds studied to date are allosterically activated by their substrate pyruvate. However, detailed kinetic studies on the enzyme from Neurospora crassa demonstrate for the first time the lack o ...201122077835
detection of antifungal compounds in polygonum ferrugineum wedd. extracts by bioassay-guided fractionation. some evidences of their mode of action.polygonum ferrugineum wedd. (polygonaceae) is used to heal infected wounds and as antiseptic, antibiotic or antifungal in the traditional argentinean medicine. the present investigation was carried out to evaluate the antifungal properties of different extracts of aerial parts of polygonum ferrugineum, in order to give support to its ethnopharmacological use and to isolate the compounds responsible for the antifungal properties. the most active compounds were tested for their capacity of produci ...201122001591
genetic dissection of parylation in the filamentous fungus neurospora crassa.parylation is a posttranslational protein modification carried out by par polymerases (parps). these enzymes function as adp-ribose transferases that add polymers of adp-ribose (par) to target proteins. parp proteins have critical functions impacting the aspects of normal human health, such as aging, as well as disease development, particularly cancer. recently, the powerful antitumor parp inhibitor olaparib was shown to be effective in blocking the progression of brca1/2-associated tumors, prom ...201121870276
[vegetable cytology. intracellular localization of transaminase alanine-glyoxylate and malate synthetase in neurospora crassa]. 20114979219
nmr structure of the c-terminal domain of a tyrosyl-trna synthetase that functions in group i intron splicing.the mitochondrial tyrosyl-trna synthetases (mt tyrrss) of pezizomycotina fungi are bifunctional proteins that aminoacylate mitochondrial trna(tyr) and are structure-stabilizing splicing cofactors for group i introns. studies with the neurospora crassa synthetase (cyt-18 protein) showed that splicing activity is dependent upon pezizomycotina-specific structural adaptations that form a distinct group i intron-binding site in the n-terminal catalytic domain. although cyt-18's c-terminal domain also ...201121438536
a widespread class of reverse transcriptase-related cellular genes.reverse transcriptases (rts) polymerize dna on rna templates. they fall into several structurally related but distinct classes and form an assemblage of rt-like enzymes that, in addition to rts, also includes certain viral rna-dependent rna polymerases (rdrp) synthesizing rna on rna templates. it is generally believed that most rt-like enzymes originate from retrotransposons or viruses and have no specific function in the host cell, with telomerases being the only notable exception. here we repo ...201121876125
crystal structure of the mid-piwi lobe of a eukaryotic argonaute protein.argonaute proteins (agos) are essential effectors in rna-mediated gene silencing pathways. they are characterized by a bilobal architecture, in which one lobe contains the n-terminal and paz domains and the other contains the mid and piwi domains. here, we present the first crystal structure of the mid-piwi lobe from a eukaryotic ago, the neurospora crassa qde-2 protein. compared to prokaryotic agos, the domain orientation is conserved, indicating a conserved mode of nucleic acid binding. the pi ...201121646546
a potassium-proton symport in neurospora crassa.combined ion flux and electrophysiological measurements have been used to characterized active transport of potassium by cells of neurospora crassa that have been moderately starved of k+ and then maintained in the presence of millimolar free calcium ions. these conditions elicit a high-affinity (k1/2 = 1-10 microm) potassium uptake system that is strongly depolarizing. current-voltage measurements have demonstrated a k+-associated inward current exceeding (at saturation) half the total current ...20113014042
systems biology of the qa gene cluster in neurospora ensemble of genetic networks that describe how the model fungal system, neurospora crassa, utilizes quinic acid (qa) as a sole carbon source has been identified previously. a genetic network for qa metabolism involves the genes, qa-1f and qa-1s, that encode a transcriptional activator and repressor, respectively and structural genes, qa-2, qa-3, qa-4, qa-x, and qa-y. by a series of 4 separate and independent, model-guided, microarray experiments a total of 50 genes are identified as qa-respon ...201121695121
array comparative genomic hybridizations: assessing the ability to recapture evolutionary relationships using an in silico approach.comparative genomic hybridization (cgh) with dna microarrays has many biological applications including surveys of copy number changes in tumorogenesis, species detection and identification, and functional genomics studies among related organisms. array cgh has also been used to infer phylogenetic relatedness among species or strains. although the use of the entire genome can be seen as a considerable advantage for use in phylogenetic analysis, few such studies have questioned the reliability of ...201121936922
characterization of cytotoxic compound from mangrove derived fungi irpex hydnoides investigate the cytotoxic activity of endophytic fungi isolated from mangrove fungi.201123569763
identification and characterization of genes required for cell-to-cell fusion in neurospora crassa.a screening procedure was used to identify cell fusion (hyphal anastomosis) mutants in the neurospora crassa single gene deletion library. mutants with alterations in 24 cell fusion genes required for cell fusion between conidial anastomosis tubes (cats) were identified and characterized. the cell fusion genes identified included 14 genes that are likely to function in signal transduction pathways needed for cell fusion to occur (mik-1, mek-1, mak-1, nrc-1, mek-2, mak-2, rac-1, pp2a, so/ham-1, h ...201121666072
fungal photobiology: a synopsis.fungi respond and adapt to many environmental signals including light. the photobiology of fungi has been extensively investigated, but in recent years the identification of the first fungal photoreceptor, wc-1 in the ascomycete neurospora crassa, and the discovery that similar photoreceptors are required for photoreception in other ascomycete, basidiomycete and zygomycete fungi has allowed the molecular characterization of light reception and the early steps of signal transduction in a number o ...201122679584
ric8 is a guanine-nucleotide exchange factor for g{alpha} subunits that regulates growth and development in neurospora crassa.heterotrimeric (aß?) g proteins are crucial components of eukaryotic signal transduction pathways. g protein coupled receptors (gpcrs) act as guanine nucleotide exchange factors (gefs) for ga subunits. recently, facilitated gdp/gtp exchange by non-gpcr gefs, such as ric8, has emerged as an important mechanism for ga regulation in animals. ric8 is present in animals and filamentous fungi, such as the model eukaryote neurospora crassa, but is absent from the genomes of baker's yeast and plants. in ...201121750256
the gene card encodes the aldehyde dehydrogenase responsible for neurosporaxanthin biosynthesis in fusarium fujikuroi.neurosporaxanthin (ß-apo-4'-carotenoic acid) biosynthesis has been studied in detail in the fungus fusarium fujikuroi. the genes and enzymes for this biosynthetic pathway are known until the last enzymatic step, the oxidation of the aldehyde group of its precursor, ß-apo-4'-carotenal. based on sequence homology to neurospora crassa ylo-1, which mediates the formation of apo-4'-lycopenoic acid from the corresponding aldehyde substrate, we cloned the card gene of f. fujikuroi and investigated the ...201121749649
Purification and characterization of heterologously expressed nitrilases from filamentous fungi.Nitrilases from Aspergillus niger CBS 513.88, A. niger K10, Gibberella moniliformis, Neurospora crassa OR74A, and Penicillium marneffei ATCC 18224 were expressed in Escherichia coli BL21-Gold (DE3) after IPTG induction. N. crassa nitrilase exhibited the highest yield of 69,000 U?L(-1) culture. Co-expression of chaperones (GroEL/ES in G. moniliformis and P. marneffei; GroEL/ES and trigger factor in N. crassa and A. niger CBS 513.88) enhanced the enzyme solubility. Specific activities of strains e ...201121892598
Expression of aspartic protease from Neurospora crassa in industrial ethanol-producing yeast and its application in ethanol production.In order to further improve the utilization rate of raw materials and increase ethanol productivity, the gene Asp, encoding aspartic protease in Neurospora crassa was cloned and expressed in industrial ethanol-producing yeast. To promote secretion of the acid protease, the gene was fused to signal sequence of the yeast a-factor gene and constitutively expressed under transcriptional control of the Saccharomyces cerevisiae PGK1 promoter. The resultant recombinant enzyme was characterized with res ...201122112824
global warming, plant paraquat resistance, and light signal transduction through nucleoside diphosphate kinase as a paradigm for increasing food supply.light signal transduction was studied in extracts of mycelia of the fungus neurospora crassa, and the third internodes of dark-grown pisum sativum cv alaska. both processes increased the phosphorylation of nucleoside diphosphate kinase (ndpk). ndpk may function as a carrier of reduction equivalents, as it binds nadh, thereby providing electrons to transform singlet oxygen to superoxide by catalases (cat). as the c-termini of ndpk interact with cat which receive singlet oxygen, emitted from photo ...201121603975
a new diet for yeast to improve biofuel 2010, our group announced the discovery of two cellodextrin transporter families from the cellulolytic fungus, neurospora crassa. furthermore, we demonstrated the utility of these transporters in the production of lignocellulosic biofuels. this discovery was made possible by a decision to systematically study cell wall degradation by n. crassa. the identified transport pathway has opened up a new way of thinking about microbial fermentation of hexoses as well as pentoses derived from plant ce ...201121637011
Xylitol production is increased by expression of codon-optimized Neurospora crassa xylose reductase gene in Candida tropicalis.Xylose reductase (XR) is the first enzyme in D: -xylose metabolism, catalyzing the reduction of D: -xylose to xylitol. Formation of XR in the yeast Candida tropicalis is significantly repressed in cells grown on medium that contains glucose as carbon and energy source, because of the repressive effect of glucose. This is one reason why glucose is not a suitable co-substrate for cell growth in industrial xylitol production. XR from the ascomycete Neurospora crassa (NcXR) has high catalytic effici ...201121922311
transgenic nicotiana tabacum plants expressing a fungal copper transporter gene show enhanced acquisition of copper.the diets of two-thirds of the world's population are deficient in one or more essential elements and one of the approaches to enhance the levels of mineral elements in food crops is by developing plants with ability to accumulate them in edible parts. besides conventional methods, transgenic technology can be used for enhancing metal acquisition in plants. copper is an essential element, which is often deficient in human diet. with the objective of developing plants with improved copper acquisi ...201121671073
[the different effects of carbon dioxide on the toxicity of silver ions for prokaryotic and eukaryotic microorganisms].the effect of carbon dioxide on survivability of bacteria escherichia coli and the germination ability ofconidia of the fungus neurospora crassa in the presence of silver nitrate was studied. it was shown that carbon dioxide increased the toxic effect of silver ions on prokaryotic cells of e. coli but did not change the survivability of spores of the eukaryote n. crassa.201121790000
mid1, a mechanosensitive calcium ion channel, affects growth, development, and ascospore discharge in the filamentous fungus gibberella zeae.the role of mid1, a stretch-activated ion channel capable of being permeated by calcium, in ascospore development and forcible discharge from asci was examined in the pathogenic fungus gibberella zeae (anamorph fusarium graminearum). the ?mid1 mutants exhibited a >12-fold reduction in ascospore discharge activity and produced predominately abnormal two-celled ascospores with constricted and fragile septae. the vegetative growth rate of the mutants was ~50% of the wild-type rate, and production o ...201121357477
solution structures of dead-box rna chaperones reveal conformational changes and nucleic acid tethering by a basic tail.the mitochondrial dead-box proteins mss116p of saccharomyces cerevisiae and cyt-19 of neurospora crassa are atp-dependent helicases that function as general rna chaperones. the helicase core of each protein precedes a c-terminal extension and a basic tail, whose structural role is unclear. here we used small-angle x-ray scattering to obtain solution structures of the full-length proteins and a series of deletion mutants. we find that the two core domains have a preferred relative orientation in ...201121746911
role of four calcium transport proteins, encoded by nca-1, nca-2, nca-3, and cax, in maintaining intracellular calcium levels in neurospora crassa.we have examined the distribution of calcium in neurospora crassa and investigated the role of four predicted calcium transport proteins. the results of cell fractionation experiments showed 4% of cellular calcium in mitochondria, approximately 11% in a dense vacuolar fraction, 40% in an insoluble form that copurifies with microsomes, and 40% in a high-speed supernatant, presumably from large vacuoles that had broken. strains lacking nca-1, a serca-type ca(2+)-atpase, or nca-3, a pmc-type ca(2+) ...201121335528
novel terpenoids of the fungus aspergillus insuetus isolated from the mediterranean sponge psammocinia sp. collected along the coast of israel.three novel meroterpenoids, insuetolides a-c (1-3) and four drimane sesquiterpenes, the new (e)-6-(4'-hydroxy-2'-butenoyl)-strobilactone a (4) and the known 2α, 9α, 11-trihydroxy-6-oxodrim-7-ene (5), strobilactone a (6) and (e,e)-6-(6',7'-dihydroxy-2',4'-octadienoyl)-strobilactone a (7), were isolated from the etoac extract of the culture medium of the marine-derived fungus aspergillus insuetus (oy-207), which was isolated from the mediterranean sponge psammocinia sp. the structures of the compo ...201121676619
functional refolding and characterization of two tom40 isoforms from human mitochondria.tom40 proteins represent an essential class of molecules which facilitate translocation of unfolded proteins from the cytosol into the mitochondrial intermembrane space. they are part of a high-molecular mass complex that forms the protein-conducting channel in outer mitochondrial membranes. this study concerns the recombinant expression, purification and folding of amino-terminally truncated variants of the two human tom40 isoforms for structural biology experiments. both cd and ftir secondary ...201121717124
regulation of stomatal tropism and infection by light in cercospora zeae-maydis: evidence for coordinated host/pathogen responses to photoperiod?cercospora zeae-maydis causes gray leaf spot of maize, which has become one of the most widespread and destructive diseases of maize in the world. c. zeae-maydis infects leaves through stomata, which is predicated on the ability of the pathogen to perceive stomata and reorient growth accordingly. in this study, the discovery that light was required for c. zeae-maydis to perceive stomata and infect leaves led to the identification of crp1, a gene encoding a putative blue-light photoreceptor homol ...201121829344
Involvement of a helix-loop-helix transcription factor CHC-1 in CO(2)-mediated conidiation suppression in Neurospora crassa.The morphological switch from vegetative growth to conidiation in filamentous fungi is highly regulated, but the understanding of the regulatory mechanisms is limited. In this study, by screening a set of knock-out mutants corresponding to 103 transcription factor encoding genes in Neurospora crassa, a mutant was found to produce abundant conidia in race tubes in which conidiation in the wild-type strain was suppressed. The corresponding gene NCU00749 encodes a protein containing a helix-loop-he ...201122001287
progressive cavitating leukoencephalopathy associated with respiratory chain complex i deficiency and a novel mutation in ndufs1.we present clinical, neuroimaging, and molecular data on the identification of a new homozygous c.1783a>g (p.thr595ala) mutation in ndufs1 in two inbred siblings with isolated complex i deficiency associated to a progressive cavitating leukoencephalopathy, a clinical and neuroradiological entity originally related to unknown defects of the mitochondrial energy metabolism. in both sibs, the muscle biopsy showed severe reduction of complex i enzyme activity, which was not obvious in fibroblasts. w ...201121203893
aspergillus nidulans transcription factor atfa interacts with the mapk saka to regulate general stress responses, development and spore functions.fungi utilize a phosphorelay system coupled to a map kinase module for sensing and processing environmental signals. in aspergillus nidulans, response regulator sska transmits osmotic and oxidative stress signals to the stress mapk (sapk) saka. using a genetic approach together with gfp tagging and molecular bifluorescence we show that saka and atf/creb transcription factor atfa define a general stress-signalling pathway that plays differential roles in oxidative stress responses during growth a ...201121320182
combined use of paracoccidioides brasiliensis recombinant rpb27 and rpb40 antigens in an enzyme-linked immunosorbent assay for immunodiagnosis of paracoccidioidomycosis.paracoccidioidomycosis (pcm) is one of the most important endemic mycoses in latin america; it's usually diagnosed by observation and/or isolation of the etiologic agent, paracoccidioides brasiliensis, as well as by a variety of immunological methods, such as complement fixation and immunodiffusion. although these approaches are useful, historically their sensitivity and specificity have often been compromised by the use of complex mixtures of undefined antigens. the use of combinations of purif ...201121354162
identification of macrodomain proteins as novel o-acetyl-adp-ribose deacetylases.sirtuins are a family of protein lysine deacetylases, which regulate gene silencing, metabolism, life span, and chromatin structure. sirtuins utilize nad(+) to deacetylate proteins, yielding o-acetyl-adp-ribose (oaadpr) as a reaction product. the macrodomain is a ubiquitous protein module known to bind adp-ribose derivatives, which diverged through evolution to support many different protein functions and pathways. the observation that some sirtuins and macrodomains are physically linked as fusi ...201121257746
heterochromatin is required for normal distribution of neurospora crassa cenh3.centromeres serve as platforms for the assembly of kinetochores and are essential for nuclear division. here we identified neurospora crassa centromeric dna by chromatin immunoprecipitation followed by high-throughput sequencing (chip-seq) of dna associated with tagged versions of the centromere foundation proteins cenh3 (cenp-a) and cen-c (cenp-c) and the kinetochore protein cen-t (cenp-t). on each chromosome we found an ∼150- to 300-kbp region of enrichment for all three proteins. these region ...201121505064
cross-talk between the cellular redox state and the circadian system in neurospora.the circadian system is composed of a number of feedback loops, and multiple feedback loops in the form of oscillators help to maintain stable rhythms. the filamentous fungus neurospora crassa exhibits a circadian rhythm during asexual spore formation (conidiation banding) and has a major feedback loop that includes the frequency (frq)/white collar (wc) -1 and -2 oscillator (fwo). a mutation in superoxide dismutase (sod)-1, an antioxidant gene, causes a robust and stable circadian rhythm compare ...201122164247
catalytic properties and classification of cellobiose dehydrogenases from ascomycetes.putative cellobiose dehydrogenase (cdh) genes are frequently discovered in various fungi by genome sequencing projects. the expression of cdh, an extracellular flavocytochrome, is well studied in white rot basidiomycetes and is attributed to extracellular lignocellulose degradation. cdh has also been reported for plant-pathogenic or saprotrophic ascomycetes, but the molecular and catalytic properties of these enzymes are currently less investigated. this study links various ascomycetous cdh gene ...201121216904
acon-3, the neurospora crassa ortholog of the developmental modifier, meda, complements the conidiation defect of the aspergillus nidulans mutant.aspergillus nidulans and neurospora crassa are ascomycetes that produce asexual spores through morphologically distinct processes. meda, a protein with unknown function, is required for normal asexual and sexual development in a. nidulans. we determined that the n. crassa ortholog of meda is acon-3, a gene required for early conidiophore development and female fertility. to test hypotheses about the evolutionary origins of asexual development in distinct fungal lineages it is important to unders ...201121220038
expression of fungal diacylglycerol acyltransferase2 genes to increase kernel oil in maize.maize (zea mays) oil has high value but is only about 4% of the grain by weight. to increase kernel oil content, fungal diacylglycerol acyltransferase2 (dgat2) genes from umbelopsis (formerly mortierella) ramanniana and neurospora crassa were introduced into maize using an embryo-enhanced promoter. the protein encoded by the n. crassa gene was longer than that of u. ramanniana. it included 353 amino acids that aligned to the u. ramanniana dgat2a protein and a 243-amino acid sequence at the amino ...201121245192
expression of cellobiose dehydrogenase from neurospora crassa in pichia pastoris and its purification and characterization.a gene encoding cellobiose dehydrogenase (cdh) from neurospora crassa strain fgsc 2489 has been cloned and expressed in the heterologous host pichia pastoris, under the control of the aox1 methanol inducible promoter. recombinant cdh without the native signal sequence and fused with a his(6)-tag (rnc-cdh1) was successfully expressed and secreted. rnc-cdh1 was produced at the level of 652 iu/l after 2 days of cultivation in the induction medium. the his(6)-tagged rnc-cdh1 was purified through a o ...201120709172
nmr solution structure of a cyanovirin homolog from wheat head blight fungus.members of the cyanovirin-n homolog (cvnh) lectin family are found in bacteria, fungi and plants. as part of our ongoing work on cvnh structure-function studies, we determined the high-resolution nmr solution structure of the homolog from the wheat head blight disease causing ascomycetous fungus gibberella zeae (or fusarium graminearum), hereafter called gzcvnh. like cyanovirin-n (cv-n), gzcvnh comprises two tandem sequence repeats and the protein sequence exhibits 30% identity with cv-n. the ov ...201121365681
inactivation of snt2, a bah/phd-containing transcription factor, impairs pathogenicity and increases autophagosome abundance in fusarium oxysporum.the soil-borne, asexual fungus fusarium oxysporum f.sp. melonis (fom) is a causal agent of muskmelon wilt disease. the current study focused on the most virulent race of fom-race 1,2. the tagged mutant d122, generated by agrobacterium tumefaciens-mediated transformation, caused the delayed appearance of initial wilt disease symptoms, as well as a 75% reduction in pathogenicity. d122 was impaired in the gene product homologous to the snt2-like transcription factor of schizosaccharomyces pombe. in ...201121535351
exploring the bzip transcription factor regulatory network in neurospora crassa.transcription factors (tfs) are key nodes of regulatory networks in eukaryotic organisms, including filamentous fungi such as neurospora crassa. the 178 predicted dna-binding tfs in n. crassa are distributed primarily among six gene families, which represent an ancient expansion in filamentous ascomycete genomes; 98 tf genes show detectable expression levels during vegetative growth of n. crassa, including 35 that show a significant difference in expression level between hyphae at the periphery ...201021081763
conserved components, but distinct mechanisms for the placement and assembly of the cell division machinery in unicellular and filamentous ascomycetes.cytokinesis is essential for cell proliferation, yet its molecular description is challenging, because >100 conserved proteins must be spatially and temporally co-ordinated. despite the high importance of a tight co-ordination of cytokinesis with chromosome and organelle segregation, the mechanism for determining the cell division plane is one of the least conserved aspects of cytokinesis in eukaryotic cells. budding and fission yeast have developed fundamentally distinct mechanisms to ensure pr ...201021091496
mips: curated databases and comprehensive secondary data resources in 2010.the munich information center for protein sequences (mips at the helmholtz center for environmental health, neuherberg, germany) has many years of experience in providing annotated collections of biological data. selected data sets of high relevance, such as model genomes, are subjected to careful manual curation, while the bulk of high-throughput data is annotated by automatic means. high-quality reference resources developed in the past and still actively maintained include saccharomyces cerev ...201021109531
archazolid a binds to the equatorial region of the c-ring of the vacuolar h+-atpase.the macrolactone archazolid is a novel, highly specific v-atpase inhibitor with an ic(50) value in the low nanomolar range. the binding site of archazolid is presumed to overlap with the binding site of the established plecomacrolide v-atpase inhibitors bafilomycin and concanamycin in subunit c of the membrane-integral v(o) complex. using a semi-synthetic derivative of archazolid for photoaffinity labeling of the v(1)v(o) holoenzyme we confirmed binding of archazolid to the v(o) subunit c. for t ...201020884613
genome-wide prediction of g protein-coupled receptors in verticillium spp.g protein-coupled receptors (gpcrs) are critical factors in regulating morphogenesis, mating, infection and virulence in fungi. in this study, various computational strategies were applied to identify gpcr-like proteins from the genomes of both verticillium dahliae and verticillium albo-atrum. the putative gpcrs were distributed over 13 classes, and significantly, three of those represented novel classes of gpcr-like proteins in fungi. the three novel gpcrs had high levels of identity to their c ...201020943146
a circadian rhythm regulating hyphal melanization in cercospora kikuchii.many metabolic and developmental processes in fungi are controlled by biological rhythms. circadian rhythms approximate a daily (24 h) cycle and have been thoroughly studied in the model fungus, neurospora crassa. however relatively few examples of true circadian rhythms have been documented among other filamentous fungi. in this study we describe a circadian rhythm underlying hyphal melanization in cercospora kikuchii, an important pathogen of soybean. after growth in light or light : dark cycl ...201020943572
cellodextrin transport in yeast for improved biofuel production.fungal degradation of plant biomass may provide insights for improving cellulosic biofuel production. we show that the model cellulolytic fungus neurospora crassa relies on a high-affinity cellodextrin transport system for rapid growth on cellulose. reconstitution of the n. crassa cellodextrin transport system in saccharomyces cerevisiae promotes efficient growth of this yeast on cellodextrins. in simultaneous saccharification and fermentation experiments, the engineered yeast strains more rapid ...201020829451
rna interference pathways in filamentous fungi.rna interference is a conserved homology-dependent post-transcriptional/transcriptional gene silencing mechanism in eukaryotes. the filamentous fungus neurospora crassa is one of the first organisms used for rnai studies. quelling and meiotic silencing by unpaired dna are two rnai-related phenomena discovered in neurospora, and their characterizations have contributed significantly to our understanding of rnai mechanisms in eukaryotes. a type of dna damage-induced small rna, microrna-like small ...201020680389
identification of pex33, a novel component of the peroxisomal docking complex in the filamentous fungus neurospora crassa.the docking complex of peroxisomal matrix protein import is composed of pex13 and pex14 in all species analyzed so far, whereas only yeast appears to possess an additional component, pex17. in this report we isolated pex14 complexes of neurospora crassa. among the complex constituents, one protein designated as pex33 possessed homology to pex14 but only in a short n-terminal domain. the pex14/pex33 interaction was verified by means of two-hybrid analysis. moreover, pex33 was shown to interact wi ...201020728240
confining euchromatin/heterochromatin territory: jumonji crosses the line.heterochromatin is typically highly condensed, gene-poor, and transcriptionally silent, whereas euchromatin is less condensed, gene-rich, and more accessible to transcription. besides acting as a graveyard for selfish mobile dna repeats, heterochromatin contributes to important biological functions, such as chromosome segregation during cell division. multiple features of heterochromatin-including the presence or absence of specific histone modifications, dna methylation, and small rnas-have bee ...201020634313
[spectrophometric analysis of volatile compounds in microorganisms].a simple modification of a spectrophometric method was proposed for the rapid detection of microorganisms based on their ability either to excrete or to absorb volatile compounds. the method provides the possibility of contactless control for bacterial growth at a concentration above 10(7) cells/ml. in addition, the method allows discriminating mutants of the fungus neurospora crassa defective in the nitrogen metabolism from the wild type strains. it is likely that nitrite reductase and nitrate ...201020586286
genome mining for the discovery of new nitrilases in filamentous fungi.purpose of work: our aim is to describe new fungal nitrilases whose sequences were published but whose catalytic properties were unknown. we adapted for expression in e. coli three of the genes and confirmed that the enzymes acted on organic nitriles. the genome mining approach was used to search for nitrilases in filamentous fungi. synthetic genes encoding nitrilases in aspergillus niger, gibberella moniliformis and neurospora crassa were expressed in escherichia coli. this is the first heterol ...201020882316
extensive and varied modifications in histone h2b of wild-type and histone deacetylase 1 mutant neurospora crassa.dna methylation is deficient in a histone deacetylase 1 (hda1) mutant (hda-1) strain of neurospora crassa with inactivated histone deacetylase 1. difference two-dimensional (2d) gels identified the primary histone deacetylase 1 target as histone h2b. acetylation was identified by lc-ms/ms at five different lysines in wild-type h2b and at 11 lysines in hda-1 h2b, suggesting neurospora h2b is a complex combination of different acetylated species. individual 2d gel spots were shifted by single lysi ...201020462202
the mitogen-activated protein kinase kinase bos5 is involved in regulating vegetative differentiation and virulence in botrytis cinerea.we present a characterization of bos5 from botrytis cinerea, a gene that encodes a mitogen-activated protein kinase kinase (mapkk), which is homologous to os-5 of neurospora crassa. the bos5 gene deletion mutant exhibited reduced vegetative growth and strongly impaired conidiation. the mutant also exhibited increased sensitivity to the dicarboximide fungicide iprodione and to osmotic stress mediated by nacl or kcl. western-blot analysis showed that the bcsak1 protein, the putative downstream com ...201020595070
characterization of a thioredoxin (bbtrx) from the entomopathogenic fungus beauveria bassiana and its expression in response to thermal stress.a thioredoxin (bbtrx) was identified from the entomopathogenic fungus beauveria bassiana. the cloned nucleotide sequence consisted of a 423-bp open reading frame encoding a 141-amino-acid thioredoxin, a 1011-bp 5' region, and a 419-bp 3' region. the deduced protein sequence of bbtrx, including a common 95-amino-acid conserved domain and a unique 46-amino-acid carboxy terminal region, was similar (≤38% identity) to that of other thioredoxins and phylogenetically closest to that from neurospora cr ...201021076484
expression of a neurospora crassa zinc transporter gene in transgenic nicotiana tabacum enhances plant zinc accumulation without co-transport of cadmium.zinc (zn) is an essential micronutrient required for growth and development of all organisms. deficiency of zn in humans is widespread, affecting 25% of world population and efforts are underway to develop crop plants with high levels of zn in their edible parts. when strategies for enhancing zn in crop plants are designed, it is essential to exclude cadmium (cd), a toxic analogue of zn. in the present work, a high affinity and high specificity zinc transporter gene (tzn1) from neurospora crassa ...201020492552
the role of paaac1 encoding a mitochondrial adp/atp carrier in the biosynthesis of extracellular glycolipids, mannosylerythritol lipids, in the basidiomycetous yeast pseudozyma antarctica.pseudozyma antarctica produces large amounts of the glycolipid biosurfactants known as mannosylerythritol lipids (mel), which show not only excellent surface-active properties but also versatile biochemical actions. a gene homologous with a mitochondrial adp/atp carrier was dominantly expressed in p. antarctica under mel-producing conditions on the basis of previous gene expression analysis. the gene encoding the mitochondrial adp/atp carrier of p. antarctica (paaac1) contained a putative open r ...201020146402
aspergillus oryzae type iii polyketide synthase csya is involved in the biosynthesis of 3,5-dihydroxybenzoic a novel superfamily of type iii polyketide synthases in microbes, four genes csya, csyb, csyc, and csyd, were found in the genome of aspergillus oryzae, an industrially important filamentous fungus. in order to analyze their functions, we carried out the overexpression of csya under the control of alpha-amylase promoter in a. oryzae and identified 3,5-dihydroxybenzoic acid (dhba) as the major product. feeding experiments using (13)c-labeled acetates confirmed that the acetate labeling pattern ...201020630753
functional conservation of mei4 for meiotic dna double-strand break formation from yeasts to mice.meiotic recombination is initiated by the programmed induction of dna double-strand breaks (dsbs) catalyzed by the evolutionarily conserved spo11 protein. studies in yeast have shown that dsb formation requires several other proteins, the role and conservation of which remain unknown. here we show that two of these saccharomyces cerevisiae proteins, mei4 and rec114, are evolutionarily conserved in most eukaryotes. mei4(-/-) mice are deficient in meiotic dsb formation, thus showing the functional ...201020551173
survey and analysis of simple sequence repeats in the laccaria bicolor genome, with development of microsatellite is becoming clear that simple sequence repeats (ssrs) play a significant role in fungal genome organization, and they are a large source of genetic markers for population genetics and meiotic maps. we identified ssrs in the laccaria bicolor genome by in silico survey and analyzed their distribution in the different genomic regions. we also compared the abundance and distribution of ssrs in l. bicolor with those of the following fungal genomes: phanerochaete chrysosporium, coprinopsis cinerea, ...201021132299
aoso protein accumulates at the septal pore in response to various stresses in the filamentous fungus aspergillus oryzae.filamentous ascomycetes form hyphal networks that are compartmentalized by septa which have a perforated pore allowing the passage of cytoplasm and organelles between adjacent hyphal compartments. thus, the septal pore may play an important role in the organized growth of multicellular organisms. upon hyphal injury, the septal pore is plugged by a wound-healing organelle, known as the woronin body, to prevent excessive cytoplasmic leakage. however, the movement of proteins towards the septal por ...201019945422
the antifungal activity of the penicillium chrysogenum protein paf disrupts calcium homeostasis in neurospora crassa.the antifungal protein paf from penicillium chrysogenum exhibits growth-inhibitory activity against a broad range of filamentous fungi. evidence from this study suggests that disruption of ca(2+) signaling/homeostasis plays an important role in the mechanistic basis of paf as a growth inhibitor. supplementation of the growth medium with high ca(2+) concentrations counteracted paf toxicity toward paf-sensitive molds. by using a transgenic neurospora crassa strain expressing codon-optimized aequor ...201020622001
the ndr kinase dbf-2 is involved in regulation of mitosis, conidial development, and glycogen metabolism in neurospora crassa.neurospora crassa dbf-2 encodes an ndr (nuclear dbf2-related) protein kinase, homologous to lats1, a core component of the hippo pathway. this pathway plays important roles in restraining cell proliferation and promoting apoptosis in differentiating cells. here, we demonstrate that dbf-2 is involved in three fundamental processes in a filamentous fungus: cell cycle regulation, glycogen biosynthesis, and conidiation. dbf-2 is predominantly localized to the nucleus, and most (approximately 60%) db ...201019966031
a comparative study of activity and apparent inhibition of fungal β-glucosidases.β-glucosidases (bgs) from aspergillus fumigates, aspergillus niger, aspergillus oryzae, chaetomium globosum, emericella nidulans, magnaporthe grisea, neurospora crassa, and penicillium brasilianum were purified to homogeneity, and analyzed by isothermal titration calorimetry with respect to their hydrolytic activity and its sensitivity to glucose (product) using cellobiose as substrate. global non-linear regression of several reactions, with or without added glucose, to a product inhibition equa ...201020677177
fungal development and the cop9 signalosome.the conserved cop9 signalosome (csn) multiprotein complex is located at the interface between cellular signaling, protein modification, life span and the development of multicellular organisms. csn is required for light-controlled responses in filamentous fungi. this includes the circadian rhythm of neurospora crassa or the repression of sexual development by light in aspergillus nidulans. in contrast to plants and animals, csn is not essential for fungal viability. therefore fungi are suitable ...201020934903
rhythmic conidiation in the blue-light fungus trichoderma pleuroticola.trichoderma species conidiate in response to blue light, however, unlike in the blue-light model fungus neurospora crassa, conidiation in trichoderma spp. has been considered to be non-circadian. in this study we uncovered evidence for circadian conidiation in trichoderma pleuroticola and identified orthologues of the key n. crassa clock components, wc-1 (blr-1) and frq.201020943132
live-cell imaging of conidial fusion in the bean pathogen, colletotrichum lindemuthianum.fusion of conidia and conidial germlings by means of conidial anastomosis tubes (cats) is a common phenomenon in filamentous fungi, including many plant pathogens. it has a number of different roles, and has been speculated to facilitate parasexual recombination and horizontal gene transfer between species. the bean pathogen colletotrichum lindemuthianum naturally undergoes cat fusion on the host surface and within asexual fruiting bodies in anthracnose lesions on its host. it has not been previ ...201020965055
cazymes analysis toolkit (cat): web service for searching and analyzing carbohydrate-active enzymes in a newly sequenced organism using cazy database.the carbohydrate-active enzyme (cazy) database provides a rich set of manually annotated enzymes that degrade, modify, or create glycosidic bonds. despite rich and invaluable information stored in the database, software tools utilizing this information for annotation of newly sequenced genomes by cazy families are limited. we have employed two annotation approaches to fill the gap between manually curated high-quality protein sequences collected in the cazy database and the growing number of oth ...201020696711
multi-targeted priming for genome-wide gene expression assays.complementary approaches to assaying global gene expression are needed to assess gene expression in regions that are poorly assayed by current methodologies. a key component of nearly all gene expression assays is the reverse transcription of transcribed sequences that has traditionally been performed by priming the poly-a tails on many of the transcribed genes in eukaryotes with oligo-dt, or by priming rna indiscriminately with random hexamers. we designed an algorithm to find common sequence m ...201020716356
genes encoding a striatin-like protein (ham-3) and a forkhead associated protein (ham-4) are required for hyphal fusion in neurospora crassa.cell-cell fusion during fertilization and between somatic cells is an integral process in eukaryotic development. in neurospora crassa, the hyphal anastomosis mutant, ham-2, fails to undergo somatic fusion. in both humans and saccharomyces cerevisiae, homologs of ham-2 are found in protein complexes that include homologs to a striatin-like protein and a forkhead-associated (fha) protein. we identified a striatin (ham-3) gene and a fha domain (ham-4) gene in n. crassa; strains containing mutation ...201020601042
involvement of a putative response regulator brrg-1 in the regulation of sporulation, sensitivity to fungicides, and osmotic stress in botrytis cinerea.the response regulator protein is a core element of two-component signaling pathway. in this study, we investigated functions of brrg-1 of botrytis cinerea, a gene that encodes a putative response regulator protein, which is homologous to rrg-1 in neurospora crassa. the brrg-1 gene deletion mutant δbrrg1-62 was unable to produce conidia. the mutant showed increased sensitivity to osmotic stress mediated by nacl and kcl, and to oxidative stress generated by h(2)o(2). additionally, the mutant was ...201021161211
structure and applications of a rhamnolipid surfactant produced in soybean oil waste.soybean oil soapstock was utilized as an alternative carbon source for the production of rhamnolipids by pseudomonas aeruginosa lbi strain. the chemical composition and properties of the rhamnolipid mixture obtained were determined to define its potential applications. the chemical characterization of the rhamnolipid has revealed the presence of ten different homologues. the monorhamnolipid rhac10c1) and the dirhamnolipid rha2c10c10 were the main components of the mixture that showed predominanc ...201019649781
purification and characterization of a low molecular weight endo-xylanase from mushroom termitomyces clypeatus.a low molecular weight endo-xylanase (ec was purified from an edible mushroom termitomyces clypeatus grown in submerged medium with oat spelt xylan. xylanase was purified to apparent homogeneity by ammonium sulfate fractionation and gel filtration chromatography. its molecular weight was determined by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 12 kda. the enzyme was found to be most active at 50 degrees c and ph 5.0, being most stab ...201019728166
camp signaling pathway controls glycogen metabolism in neurospora crassa by regulating the glycogen synthase gene expression and phosphorylation.the camp-pka signaling pathway plays an important role in many biological processes including glycogen metabolism. in this work we investigated its role in the neurospora crassa glycogen metabolism control using mutant strains affected in components of the pathway, the cr-1 strain deficient in adenylyl cyclase activity therefore has the pka pathway not active, and the mcb strain a temperature-sensitive mutant defective in the regulatory subunit of pka therefore is a strain with constitutively ac ...201019883780
the s helix mediates signal transmission as a hamp domain coiled-coil extension in the narx nitrate sensor from escherichia coli the nitrate-responsive, homodimeric narx sensor, two cytoplasmic membrane alpha-helices delimit the periplasmic ligand-binding domain. the hamp domain, a four-helix parallel coiled-coil built from two alpha-helices (hd1 and hd2), immediately follows the second transmembrane helix. previous computational studies identified a likely coiled-coil-forming alpha-helix, the signaling helix (s helix), in a range of signaling proteins, including eucaryal receptor guanylyl cyclases, but its function re ...201019966007
expression of ribonuclease a and ribonuclease n1 in the filamentous fungus neurospora this study, we investigated the ability of the fungus neurospora crassa to produce and secrete two ribonucleases: the heterologous bovine rnase a and the endogenous rnase n(1). a set of expression vectors was constructed, each consisting of an rnase a open reading frame under the control of a specific promoter and each with a specific terminator. n. crassa transformants were analyzed at the transcriptional and protein levels. irrespective of the promoter used, all transformants showed an rnas ...201019662399
the mitochondrial porin, vdac, has retained the ability to be assembled in the bacterial outer membrane.beta-barrel proteins are present in the outer membranes (oms) of gram-negative bacteria, mitochondria, and chloroplasts. their assembly requires a machinery of which the central component, called omp85 (bama) in bacteria and tob55 (sam50) in mitochondria, is evolutionarily conserved. an open question is whether the signals in beta-barrel om proteins required for assembly via this multicomponent machinery are also conserved. to address this question, we have expressed in escherichia coli the mito ...201019959601
high-throughput construction of gene deletion cassettes for generation of neurospora crassa knockout strains.the availability of complete genome sequences for a number of biologically important fungi has become an important resource for fungal research communities. however, the functions of many open reading frames (orfs) identified through annotation of whole genome sequences have yet to be determined. the disruption of orfs is a practical method for loss-of-function gene analyses in fungi that are amenable to transformation. unfortunately, the construction of knockout cassettes using traditional dige ...201020238259
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