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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
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
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
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
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
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
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
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
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
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
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
[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
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
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
circadian timekeeping in neurospora crassa and synechococcus elongatus.at 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
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
a new diet for yeast to improve biofuel production.in 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
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
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
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
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
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
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
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
AreA controls nitrogen source utilisation during both growth programs of the dimorphic fungus Penicillium marneffei.The opportunistic pathogen Penicillium marneffei displays a temperature-dependent dimorphic switching program with saprophytic hyphal growth at 25 °C and yeast growth at 37 °C. The areA gene of P. marneffei has been isolated and found to be required for the utilisation of nonpreferred nitrogen sources during both growth programs of P. marneffei, albeit to differing degrees. Based on this functional characterisation and high degree of sequence conservation with other fungal GATA factors, P. marne ...201222208609
the neurospora crassa os mapk pathway-activated transcription factor asl-1 contributes to circadian rhythms in pathway responsive clock-controlled genes.the os-pathway mitogen-activated protein kinase (mapk) cascade of neurospora crassa is responsible for adaptation to osmotic stress. activation of the mapk, os-2, leads to the transcriptional induction of many genes involved in the osmotic stress response. we previously demonstrated that there is a circadian rhythm in the phosphorylation of os-2 under constant non-stress inducing conditions. additionally, several osmotic stress-induced genes are known to be regulated by the circadian clock. ther ...201222240319
biological activities of extracts from chenopodium ambrosioides lineu and kielmeyera neglecta saddi.chenopodium ambrosioides and kielmeyera neglecta are plants traditionally used in brazil to treat various infectious diseases. the study of the biological activities of these plants is of great importance for the detection of biologically active compounds.201222839690
prp8 inteins in species of the genus botrytis and other ascomycetes.the mobile elements termed inteins have a sporadic distribution in microorganisms. it is unclear how these elements are maintained. inteins are intervening protein sequences that autocatalytically excise themselves from a precursor. excision is a post-translational process referred to as 'protein splicing' in which the sequences flanking the intein are ligated, reforming the mature host protein. some inteins contain a homing endonuclease domain (heg) that is proposed to facilitate propagation of ...201222285471
transgene induced co-suppression during vegetative growth in cryptococcus neoformans.introduction of dna sequences into the genome often results in homology-dependent gene silencing in organisms as diverse as plants, fungi, flies, nematodes, and mammals. we previously showed in cryptococcus neoformans that a repeat transgene array can induce gene silencing at a high frequency during mating (∼50%), but at a much lower frequency during vegetative growth (∼0.2%). here we report a robust asexual co-suppression phenomenon triggered by the introduction of a cpa1::ade2 transgene. multi ...201222916030
ambient ph controls glycogen levels by regulating glycogen synthase gene expression in neurospora crassa. new insights into the ph signaling pathway.glycogen is a polysaccharide widely distributed in microorganisms and animal cells and its metabolism is under intricate regulation. its accumulation in a specific situation results from the balance between glycogen synthase and glycogen phosphorylase activities that control synthesis and degradation, respectively. these enzymes are highly regulated at transcriptional and post-translational levels. the existence of a dna motif for the aspergillus nidulans ph responsive transcription factor pacc ...201222952943
the carnitine biosynthetic pathway in arabidopsis thaliana shares similar features with the pathway of mammals and fungi.carnitine is an essential quaternary ammonium amino acid that occurs in the microbial, plant and animal kingdoms. the role and synthesis of this compound are very well documented in bacteria, fungi and mammals. on the contrary, although the presence of carnitine in plant tissue has been reported four decades ago and information about its biological implication are available, nothing is known about its synthesis in plants. we designed experiments to determine if the carnitine biosynthetic pathway ...201222922110
fungal catalases: function, phylogenetic origin and structure.most fungi have several monofunctional heme-catalases. filamentous ascomycetes (pezizomycotina) have two types of large-size subunit catalases (l1 and l2). l2-type are usually induced by different stressors and are extracellular enzymes; those from the l1-type are not inducible and accumulate in asexual spores. l2 catalases are important for growth and the start of cell differentiation, while l1 are required for spore germination. in addition, pezizomycetes have one to four small-size subunit ca ...201222698962
a novel activity for fungal nitronate monooxygenase: detoxification of the metabolic inhibitor propionate-3-nitronate.nitronate monooxygenase (nmo; e.c. 1.13.12.16) oxidizes alkyl nitronates to aldehydes and nitrite. although the biochemistry of the enzyme from fungal sources has been studied extensively, the physiological role is unknown. the ability of nmo to detoxify propionate-3-nitronate was tested by measuring growth of recombinant escherichia coli containing the gene encoding for the enzyme in either the absence or presence of the nitronate and its conjugate acid 3-nitropropionate. the mixture propionate ...201222464989
a novel biochemical route for fuels and chemicals production from cellulosic biomass.the conventional biochemical platform featuring enzymatic hydrolysis involves five key steps: pretreatment, cellulase production, enzymatic hydrolysis, fermentation, and product recovery. sugars are produced as reactive intermediates for subsequent fermentation to fuels and chemicals. herein, an alternative biochemical route is proposed. pretreatment, enzymatic hydrolysis and cellulase production is consolidated into one single step, referred to as consolidated aerobic processing, and sugar aldo ...201222384058
small rna isolation and library construction for expression profiling of small rnas from neurospora and fusarium using illumina high-throughput deep sequencing.due to crucial roles in gene regulation, noncoding small rnas (smrnas) of 20-30 nucleotides (nt) have been intensively studied in mammals and plants, and are known to be implicated in significant diseases and metabolic disorders. elucidation of biogenesis mechanisms and functional characterization of smrnas are often achieved using tools, such as separation of small-sized rna and high-throughput sequencing. although rna interference pathways such as quelling and meiotic silencing have been well ...201222589132
gliotoxin effects on fungal growth: mechanisms and exploitation.although initially investigated for its antifungal properties, little is actually known about the effect of gliotoxin on aspergillus fumigatus and other fungi. we have observed that exposure of a. fumigatus to exogenous gliotoxin (14 μg/ml), under gliotoxin-limited growth conditions, results in significant alteration of the expression of 27 proteins (up- and down-regulated >1.9-fold; p<0.05) including de novo expression of cu, zn superoxide dismutase, up-regulated allergen asp f3 expression and ...201222405895
characterization of the insertase for β-barrel proteins of the outer mitochondrial membrane.the tob-sam complex is an essential component of the mitochondrial outer membrane that mediates the insertion of β-barrel precursor proteins into the membrane. we report here its isolation and determine its size, composition, and structural organization. the complex from neurospora crassa was composed of tob55-sam50, tob38-sam35, and tob37-sam37 in a stoichiometry of 1:1:1 and had a molecular mass of 140 kd. a very minor fraction of the purified complex was associated with one mdm10 protein. usi ...201223128244
the transcriptional response to the inactivation of the pampk1 and pampk2 map kinase pathways in podospora anserina.transcription pattern during mycelium growth of podospora anserina was assayed by microarray analysis in wild type and in mutants affected in the map kinase genes pampk1 and pampk2 and in the nadph oxidase gene panox1. 15% of the genes have their expression modified by a factor two or more as growth proceeds in wild type. the genes whose expression is modified during growth in p. anserina are either not conserved or differently regulated in neurospora crassa and aspergillus niger, two fungi for ...201222721649
improved gene targeting frequency in marine-derived filamentous fungus aspergillus glaucus by disrupting ligd.filamentous fungi from the marine environment have shown great potential as cell factories for the production of pharmacologically active metabolites, but extremely low frequency of homologous recombination brings difficulty to further molecular biology studies. to bypass this problem and develop a highly efficient gene targeting system in marine-derived filamentous fungus aspergillus glaucus, ligd, a homolog of neurospora crassa mus-53 which is considered to play a significant role in nonhomolo ...201222562375
genetic control of asexual sporulation in filamentous fungi.asexual sporulation (conidiation) in the ascomycetous filamentous fungi involves the formation of conidia, formed on specialized structures called conidiophores. conidiation in filamentous fungi involves many common themes including spatial and temporal regulation of gene expression, specialized cellular differentiation, intra-/inter-cellular communications, and response to environmental factors. the commencement, progression and completion of conidiation are regulated by multiple positive and n ...201223092920
high abundance of serine/threonine-rich regions predicted to be hyper-o-glycosylated in the secretory proteins coded by eight fungal genomes.o-glycosylation of secretory proteins has been found to be an important factor in fungal biology and virulence. it consists in the addition of short glycosidic chains to ser or thr residues in the protein backbone via o-glycosidic bonds. secretory proteins in fungi frequently display ser/thr rich regions that could be sites of extensive o-glycosylation. we have analyzed in silico the complete sets of putatively secretory proteins coded by eight fungal genomes (botrytis cinerea, magnaporthe grise ...201222994653
conserved and essential transcription factors for cellulase gene expression in ascomycete fungi.rational engineering of filamentous fungi for improved cellulase production is hampered by our incomplete knowledge of transcriptional regulatory networks. we therefore used the model filamentous fungus neurospora crassa to search for uncharacterized transcription factors associated with cellulose deconstruction. a screen of a n. crassa transcription factor deletion collection identified two uncharacterized zinc binuclear cluster transcription factors (clr-1 and clr-2) that were required for gro ...201222532664
genomic characteristics comparisons of 12 food-related filamentous fungi in trna gene set, codon usage and amino acid composition.filamentous fungi are widely exploited in food industry due to their abilities to secrete large amounts of enzymes and metabolites. the recent availability of fungal genome sequences has provided an opportunity to explore the genomic characteristics of these food-related filamentous fungi. in this paper, we selected 12 representative filamentous fungi in the areas of food processing and safety, which were aspergillus clavatus, a. flavus, a. fumigatus, a. nidulans, a. niger, a. oryzae, a. terreus ...201222305983
yeast adaptor protein, nbp2p, is conserved regulator of fungal ptc1p phosphatases and is involved in multiple signaling pathways.nbp2p is an src homology 3 (sh3) domain-containing yeast protein that is involved in a variety of cellular processes. this small adaptor protein binds to a number of different proteins through its sh3 domain, and a region n-terminal to the sh3 domain binds to the protein phosphatase, ptc1p. despite its involvement in a large number of physical and genetic interactions, the only well characterized function of nbp2p is to recruit ptc1p to the high osmolarity glycerol pathway, which results in down ...201222570491
enforcing silencing: dynamic hp1 complexes in neurospora.analysis of the neurospora crassa chromodomain protein cdp-2, a component of a newly characterized hp1-containing complex, reveals a second gene-silencing mechanism and provides insights into the dynamic nature of chromatin domains that possess shared components.201222551706
the arginine attenuator peptide interferes with the ribosome peptidyl transferase center.the fungal arginine attenuator peptide (aap) is encoded by a regulatory upstream open reading frame (uorf). the aap acts as a nascent peptide within the ribosome tunnel to stall translation in response to arginine (arg). the effect of aap and arg on ribosome peptidyl transferase center (ptc) function was analyzed in neurospora crassa and wheat germ translation extracts using the transfer of nascent aap to puromycin as an assay. in the presence of a high concentration of arg, the wild-type aap in ...201222508989
self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin eas.the hydrophobin eas from the fungus neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. self-assembly of eas into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. we have used site-directed mutagenesis and peptide experiments to identify the region of eas that drives intermolecular association and formation of the cross-β rodlet stru ...201222308366
[analysis of several key problems about the two linkage gene mapping of neurospora crassa in genetics teaching].ordered tetrad analysis is important content in the genetics teaching. in particular, the two linkage gene mapping is not only a key point, but also a difficult one. how to explain the content better is a hard nut for the many genetics teachers or editors of the teaching material to crack. here, based on teaching practice of many years we summarized several key problems, which are difficult to understand by students and frequently neglected by the teachers and the editors of genetics. furthermor ...201222306881
a fluorescence-based supramolecular tandem assay for monitoring lysine methyltransferase activity in homogeneous solution.the demand for practical and convenient enzyme assays for histone lysine methyltransferases (hkmts) emerges along with the rapid development of this young class of enzymes. a supramolecular reporter pair composed of p-sulfonatocalix[4]arene (cx4) and the fluorescent dye lucigenin (lcg) has been used to monitor enzymatic trimethylation of lysine residues in peptide substrates. the assay affords a switch-on fluorescence response and operates in a continuous, real-time, and label-free fashion. the ...201222367964
roles for receptors, pheromones, g proteins, and mating type genes during sexual reproduction in neurospora crassa.here we characterize the relationship between the pre-2 pheromone receptor and its ligand, ccg-4, and the general requirements for receptors, pheromones, g proteins, and mating type genes during fusion of opposite mating-type cells and sexual sporulation in the multicellular fungus neurospora crassa. pre-2 is highly expressed in mat a cells and is localized in male and female reproductive structures. δpre-2 mat a females do not respond chemotropically to mat a males (conidia) or form mature frui ...201222298702
comparative live-cell imaging analyses of spa-2, bud-6 and bni-1 in neurospora crassa reveal novel features of the filamentous fungal polarisome.a key multiprotein complex involved in regulating the actin cytoskeleton and secretory machinery required for polarized growth in fungi, is the polarisome. recognized core constituents in budding yeast are the proteins spa2, pea2, aip3/bud6, and the key effector bni1. multicellular fungi display a more complex polarized morphogenesis than yeasts, suggesting that the filamentous fungal polarisome might fulfill additional functions. in this study, we compared the subcellular organization and dynam ...201222291944
excitable behavior can explain the "ping-pong" mode of communication between cells using the same chemoattractant.here we elucidate a paradox: how a single chemoattractant-receptor system in two individuals is used for communication despite the seeming inevitability of self-excitation. in the filamentous fungus neurospora crassa, genetically identical cells that produce the same chemoattractant fuse via the homing of individual cell protrusions toward each other. this is achieved via a recently described "ping-pong" pulsatile communication. using a generic activator-inhibitor model of excitable behavior, we ...201222271443
cooperation among germinating spores facilitates the growth of the fungus, neurospora crassa.fusions between individuals are a common feature of organisms with modular, indeterminate life forms, including plants, marine invertebrates and fungi. the consequences of fusion for an individual fungus are poorly understood. we used wild-type and fusion mutant strains of the genetic model neurospora crassa to chronicle the fitness in two different laboratory habitats, and in each experiment started colonies from multiple different densities of asexual spores. on round petri dishes, fusion enab ...201222258449
genome-wide selection on codon usage at the population level in the fungal model organism neurospora crassa.many organisms exhibit biased codon usage in their genome, including the fungal model organism neurospora crassa. the preferential use of subset of synonymous codons (optimal codons) at the macroevolutionary level is believed to result from a history of selection to promote translational efficiency. at present, few data are available about selection on optimal codons at the microevolutionary scale, that is, at the population level. herein, we conducted a large-scale assessment of codon mutations ...201222334579
selection of a dna barcode for nectriaceae from fungal whole-genomes.a dna barcode is a short segment of sequence that is able to distinguish species. a barcode must ideally contain enough variation to distinguish every individual species and be easily obtained. fungi of nectriaceae are economically important and show high species diversity. to establish a standard dna barcode for this group of fungi, the genomes of neurospora crassa and 30 other filamentous fungi were compared. the expect value was treated as a criterion to recognize homologous sequences. four c ...201222314494
sex-specific gene expression during asexual development of neurospora crassa.the impact of loci that determine sexual identity upon the asexual, dominant stage of fungal life history has been well studied. to investigate their impact, expression differences between strains of different mating type during asexual development were assayed, with rna sampled from otherwise largely isogenic mat a and mat a strains of neurospora crassa at early, middle, and late clonal stages of development. we observed significant differences in overall gene expression between mating types ac ...201222626843
the functions of grainy head-like proteins in animals and fungi and the evolution of apical extracellular barriers.the grainy head (grh) family of transcription factors are crucial for the development and repair of epidermal barriers in all animals in which they have been studied. this is a high-level functional conservation, as the known structural and enzymatic genes regulated by grh proteins differ between species depending on the type of epidermal barrier being formed. interestingly, members of the cp2 superfamily of transcription factors, which encompasses the grh and lsf families in animals, are also f ...201222590528
neurospora cop9 signalosome integrity plays major roles for hyphal growth, conidial development, and circadian function.the cop9 signalosome (csn) is a highly conserved multifunctional complex that has two major biochemical roles: cleaving nedd8 from cullin proteins and maintaining the stability of crl components. we used mutation analysis to confirm that the jamm domain of the csn-5 subunit is responsible for nedd8 cleavage from cullin proteins in neurospora crassa. point mutations of key residues in the metal-binding motif (ex(n)hxhx(10)d) of the csn-5 jamm domain disrupted csn deneddylation activity without in ...201222589747
on-target labeling of intracellular metabolites combined with chemical mapping of individual hyphae revealing cytoplasmic relocation of isotopologues.a microscale analytical platform integrating microbial cell culture, isotopic labeling, along with visual and mass spectrometric imaging with single-cell resolution has been developed and applied in the monitoring of cellular metabolism in fungal mycelium. the method implements open chips with a two-dimensional surface pattern composed of hydrophobic and hydrophilic zones. two hydrophilic islands are used as medium reservoirs, while the hydrophobic area constitutes the support for the growing ae ...201222583035
production of platinum nanoparticles and nanoaggregates using neurospora crassa.fungal biomass and fungal extract of the nonpathogenic fungus neurospora crassa were successfully used as reducing agents for the biosynthesis of platinum nanoparticles (ptnps). the experiment was carried out by exposing the fungal biomass or the fungal extract to a 0.001 m precursor solution of hexachloroplatinic(iv) acid (h2ptcl6). a change of color of the biomass from pale yellow to dark brown was the first indication of possible formation of ptnps by the fungus. subsequent analyses confirmed ...201222580320
controlling gene expression with the q repressible binary expression system in caenorhabditis elegans.we established a transcription-based binary gene expression system in caenorhabditis elegans using the recently developed q system. this system, derived from genes in neurospora crassa, uses the transcriptional activator qf to induce the expression of target genes. activation can be efficiently suppressed by the transcriptional repressor qs, and suppression can be relieved by the nontoxic small molecule quinic acid. we used qf, qs and quinic acid to achieve temporal and spatial control of transg ...201222406855
neurospora mrc1 homologue is involved in replication stability and is required for normal cell growth and chromosome integrity in mus-9 and mus-21 mutants.stalled replication forks easily collapse and such structures can induce dna strand breaks or toxic recombination products. therefore, factors involved in stabilization of replication should be important for genome integrity. in our previous study, loss of both atm and atr homologues was shown to cause growth defects and chromosome instability in neurospora crassa. to elucidate the relationships between these defects and replication instability, we focused on one of the viable replication factor ...201222395065
temperature-sensitive and circadian oscillators of neurospora crassa share components.in neurospora crassa, the interactions between products of the frequency (frq), frequency-interacting rna helicase (frh), white collar-1 (wc-1), and white collar-2 (wc-2) genes establish a molecular circadian clockwork, called the frq-wc-oscillator (fwo), which is required for the generation of molecular and overt circadian rhythmicity. in strains carrying nonfunctional frq alleles, circadian rhythms in asexual spore development (conidiation) are abolished in constant conditions, yet conidiation ...201222367035
high-resolution spatiotemporal analysis of gene expression in real time: in vivo analysis of circadian rhythms in neurospora crassa using a frequency-luciferase translational reporter.the pacemaker of the neurospora circadian clock is composed of a transcriptional-translational feedback loop that has been intensively studied during the last two decades. invaluable information has been derived from measuring the expression of the central clock component frequency (frq) under liquid culture conditions. direct analyses of frq mrna and protein levels on solid media - where overt circadian rhythms are normally visualized - have not been trivial due to technical issues. nevertheles ...201222695169
coronin is a component of the endocytic collar of hyphae of neurospora crassa and is necessary for normal growth and morphogenesis.coronin plays a major role in the organization and dynamics of actin in yeast. to investigate the role of coronin in a filamentous fungus (neurospora crassa), we examined its subcellular localization using fluorescent proteins and the phenotypic consequences of coronin gene (crn-1) deletion in hyphal morphogenesis, spitzenkörper behavior and endocytosis. coronin-gfp was localized in patches, forming a subapical collar near the hyphal apex; significantly, it was absent from the apex. the subapica ...201222693603
organic acid excretion in penicillium ochrochloron increases with ambient ph.despite being of high biotechnological relevance, many aspects of organic acid excretion in filamentous fungi like the influence of ambient ph are still insufficiently understood. while the excretion of an individual organic acid may peak at a certain ph value, the few available studies investigating a broader range of organic acids indicate that total organic acid excretion rises with increasing external ph. we hypothesized that this phenomenon might be a general response of filamentous fungi t ...201222493592
induction of lignocellulose-degrading enzymes in neurospora crassa by cellodextrins.neurospora crassa colonizes burnt grasslands in the wild and metabolizes both cellulose and hemicellulose from plant cell walls. when switched from a favored carbon source such as sucrose to cellulose, n. crassa dramatically upregulates expression and secretion of a wide variety of genes encoding lignocellulolytic enzymes. however, the means by which n. crassa and other filamentous fungi sense the presence of cellulose in the environment remains unclear. here, we show that an n. crassa mutant ca ...201222474347
use of fluorescent protein to analyse recombination at three loci in neurospora crassa.we have inserted a histone h1-gfp fusion gene adjacent to three loci on different chromosomes of neurospora crassa and made mating pairs in which a wild type version of gfp is crossed to one with a mutation in the 5' end of gfp. the loci are his-3, am and his-5, chosen because recombination mechanisms appear to differ between his-3 and am, and because crossing over adjacent to his-5, like his-3, is regulated by rec-2. at his-3, the frequencies of crossing over between gfp and the centromere and ...201222691725
analyses of dynein heavy chain mutations reveal complex interactions between dynein motor domains and cellular dynein functions.cytoplasmic dynein transports cargoes for a variety of crucial cellular functions. however, since dynein is essential in most eukaryotic organisms, the in-depth study of the cellular function of dynein via genetic analysis of dynein mutations has not been practical. here, we identify and characterize 34 different dynein heavy chain mutations using a genetic screen of the ascomycete fungus neurospora crassa, in which dynein is nonessential. interestingly, our studies show that these mutations seg ...201222649085
differential impact of nutrition on developmental and metabolic gene expression during fruiting body development in neurospora crassa.fungal fruiting body size and form are influenced by the ecology of the species, including diverse environmental stimuli. accordingly, nutritional resources available to the fungus during development can be vital to successful production of fruiting bodies. to investigate the effect of nutrition, perithecial development of neurospora crassa was induced on two different media, a chemically sparsely nutritive synthetic crossing medium (scm) and a natural carrot agar (ca). protoperithecia were coll ...201222469835
unravelling the molecular basis for light modulated cellulase gene expression - the role of photoreceptors in neurospora crassa.light represents an important environmental cue, which exerts considerable influence on the metabolism of fungi. studies with the biotechnological fungal workhorse trichoderma reesei (hypocrea jecorina) have revealed an interconnection between transcriptional regulation of cellulolytic enzymes and the light response. neurospora crassa has been used as a model organism to study light and circadian rhythm biology. we therefore investigated whether light also regulates transcriptional regulation of ...201222462823
analysis of the role of transcription factor vad-5 in conidiation of neurospora crassa.conidiation is the major mode of reproduction in many filamentous fungi. the neurospora crassa gene vad-5, which encodes a gal4-like zn2cys6 transcription factor, was suggested to contribute to conidiation in a previous study using a knockout mutant. in this study, we confirmed the positive contribution of vad-5 to conidiation by gene complementation. to understand the role of vad-5 in conidiation, transcriptomic profiles generated by digital gene expression profiling from the vad-5 deletion mut ...201222445960
concentration-dependent mechanisms of cell penetration and killing by the de novo designed antifungal hexapeptide paf26.recent evidence indicates that antimicrobial peptides can kill microbes in more complex ways than just by membrane permeabilization. in this study, the mechanism of internalization of the de novo designed cationic hexapeptide paf26 has been characterized in detail using neurospora crassa. live-cell imaging of fluorescently labelled paf26, organelle probes and mutants indicate that the peptide is endocytically internalized at low fungicidal concentrations (2.0-5 µm). at these concentrations, paf2 ...201222646057
role of fig1, a component of the low-affinity calcium uptake system, in growth and sexual development of filamentous fungi.the function of fig1, a transmembrane protein of the low-affinity calcium uptake system (lacs) in fungi, was examined for its role in the growth and development of the plant pathogen fusarium graminearum. the δfig1 mutants failed to produce mature perithecia, and sexual development was halted prior to the formation of perithecium initials. the loss of fig1 function also resulted in a reduced vegetative growth rate. macroconidium production was reduced 70-fold in the δfig1 mutants compared to the ...201222635922
investigation of the ph-dependent electron transfer mechanism of ascomycetous class ii cellobiose dehydrogenases on electrodes.cellobiose dehydrogenase (cdh) is capable of direct electron transfer (det) on various carbon and thiol-modified gold electrodes. as a result, these systems have been utilized as biocatalyst in biosensors and biofuel cell anodes. class i cdhs, from basidiomycetous fungi, are highly specific to cellulose or lactose, and det is only observed at ph values below 5.5. to extend the applicability of cdh-based electrodes, the catalytic properties and the behavior on electrode surfaces of ascomycetous c ...201222471986
characterization of apoptosis-related oxidoreductases from neurospora crassa.the genome from neurospora crassa presented three open reading frames homologous to the genes coding for human aif and amid proteins, which are flavoproteins with oxidoreductase activities implicated in caspase-independent apoptosis. to investigate the role of these proteins, namely within the mitochondrial respiratory chain, we studied their cellular localization and characterized the respective null mutant strains. efficiency of the respiratory chain was analyzed by oxygen consumption studies ...201222470547
hydrophobic motif phosphorylation coordinates activity and polar localization of the neurospora crassa nuclear dbf2-related kinase cot1.nuclear dbf2p-related (ndr) kinases and associated proteins are recognized as a conserved network that regulates eukaryotic cell polarity. ndr kinases require association with mob adaptor proteins and phosphorylation of two conserved residues in the activation segment and hydrophobic motif for activity and function. we demonstrate that the neurospora crassa ndr kinase cot1 forms inactive dimers via a conserved n-terminal extension, which is also required for the interaction of the kinase with mo ...201222451488
deciphering transcriptional regulatory mechanisms associated with hemicellulose degradation in neurospora crassa.hemicellulose, the second most abundant plant biomass fraction after cellulose, is widely viewed as a potential substrate for the production of liquid fuels and other value-added materials. degradation of hemicellulose by filamentous fungi requires production of many different enzymes, which are induced by biopolymers or its derivatives and regulated mainly at the transcriptional level through transcription factors (tfs). neurospora crassa, a model filamentous fungus, expresses and secretes enzy ...201222345350
electrical phenotypes of calcium transport mutant strains of a filamentous fungus, neurospora crassa.we characterized the electrical phenotypes of mutants with mutations in genes encoding calcium transporters-a mechanosensitive channel homolog (mscs), a ca(2+)/h(+) exchange protein (cax), and ca(2+)-atpases (nca-1, nca-2, nca-3)-as well as those of double mutants (the nca-2 cax, nca-2 nca-3, and nca-3 cax mutants). the electrical characterization used dual impalements to obtain cable-corrected current-voltage measurements. only two types of mutants (the mscs mutant; the nca-2 mutant and nca-2-c ...201222408225
the essential phosphoinositide kinase mss-4 is required for polar hyphal morphogenesis, localizing to sites of growth and cell fusion in neurospora crassa.fungal hyphae and plant pollen tubes are among the most highly polarized cells known and pose extraordinary requirements on their cell polarity machinery. cellular morphogenesis is driven through the phospholipid-dependent organization at the apical plasma membrane. we characterized the contribution of phosphoinositides (pis) in hyphal growth of the filamentous ascomycete neurospora crassa. mss-4 is an essential gene and its deletion resulted in spherically growing cells that ultimately lyse. tw ...201223272106
the guanine nucleotide exchange factor ric8 regulates conidial germination through gα proteins in neurospora crassa.heterotrimeric g protein signaling is essential for normal hyphal growth in the filamentous fungus neurospora crassa. we have previously demonstrated that the non-receptor guanine nucleotide exchange factor ric8 acts upstream of the gα proteins gna-1 and gna-3 to regulate hyphal extension. here we demonstrate that regulation of hyphal extension results at least in part, from an important role in control of asexual spore (conidia) germination. loss of gna-3 leads to a drastic reduction in conidia ...201223118921
production of four neurospora crassa lytic polysaccharide monooxygenases in pichia pastoris monitored by a fluorimetric assay. 201223102010
the neurospora crassa chr-1 gene is up-regulated by chromate and its encoded chr-1 protein causes chromate sensitivity and chromium accumulation.the chra membrane protein belongs to the chr superfamily of chromate ion transporters, which includes homologues from bacteria, archaea and eukaryotes. bacterial chra homologues confer chromate resistance by exporting chromate ions from the cell's cytoplasm. the neurospora crassa strain 74-a chr-1 gene encodes a putative chr-1 protein of 507 amino acid residues, which belongs to the chr superfamily. rt-pcr assays showed that expression of the chr-1 gene was up-regulated by chromate exposure of n ...201223085746
light-inducible system for tunable protein expression in neurospora crassa.filamentous fungi are important model systems for understanding eukaryotic cellular processes, including the study of protein expression. a salient feature of fungi is the ability of the protein-processing machinery to perform all of the extensive posttranslational modifications needed in the complex world of eukaryotic organisms, making them great hosts for production of eukaryotic proteins. in the model organism neurospora crassa, several regulatable promoters have been used for heterologous g ...201223050231
blue light acts as a double-edged sword in regulating sexual development of hypocrea jecorina (trichoderma reesei).the industrially important cellulolytic filamentous fungus trichoderma reesei is the anamorph of the pantropical ascomycete hypocrea jecorina. h. jecorina cbs999.97 strain undergoes a heterothallic reproductive cycle, and the mating yields fertilized perithecia imbedded in stromata. asci in the perithecia contain 16 linearly arranged ascospores. here, we investigated h. jecorina sexual development under different light regimes, and found that visible light was dispensable for sexual development ...201223028710
the ndr kinase scaffold hym1/mo25 is essential for mak2 map kinase signaling in neurospora crassa.cell communication is essential for eukaryotic development, but our knowledge of molecules and mechanisms required for intercellular communication is fragmentary. in particular, the connection between signal sensing and regulation of cell polarity is poorly understood. in the filamentous ascomycete neurospora crassa, germinating spores mutually attract each other and subsequently fuse. during these tropic interactions, the two communicating cells rapidly alternate between two different physiolog ...201223028357
the saccharomyces cerevisiae bem1 homologue in neurospora crassa promotes co-ordinated cell behaviour resulting in cell fusion.directed growth or movement is a common feature of microbial development and propagation. in polar growing filamentous fungi, directed growth requires the interaction of signal sensing machineries with factors controlling polarity and cell tip extension. in neurospora crassa an unusual mode of cell-cell signalling mediates mutual attraction of germinating spores, which subsequently fuse. during directed growth of the two fusion partners, the cells co-ordinately alternate between two physiologica ...201222906237
genetic and physical interactions between gα subunits and components of the gβγ dimer of heterotrimeric g proteins in neurospora crassa.heterotrimeric g proteins are critical regulators of growth and asexual and sexual development in the filamentous fungus neurospora crassa. three gα subunits (gna-1, gna-2, and gna-3), one gβ subunit (gnb-1), and one gγ subunit (gng-1) have been functionally characterized, but genetic epistasis relationships between gβ and gα subunit genes have not been determined. physical association between gnb-1 and flag-tagged gng-1 has been previously demonstrated by coimmunoprecipitation, but knowledge of ...201222903975
diversification of a protein kinase cascade: ime-2 is involved in nonself recognition and programmed cell death in neurospora crassa.kinase cascades and the modification of proteins by phosphorylation are major mechanisms for cell signaling and communication, and evolution of these signaling pathways can contribute to new developmental or environmental response pathways. the saccharomyces cerevisiae kinase ime2 has been well characterized for its role in meiosis. however, recent studies have revealed alternative functions for ime2 in both s. cerevisiae and other fungi. in the filamentous fungus neurospora crassa, the ime2 hom ...201222813893
graphite electrodes modified with neurospora crassa cellobiose dehydrogenase: comparative electrochemical characterization under direct and mediated electron transfer.the electrochemical characterization of a class ii cellobiose dehydrogenase (cdh), from the ascomycete fungus neurospora crassa, adsorbed on graphite (g), was performed in regard to direct (det) and mediated electron transfer (met). the effects of the applied potential, mediator (1,4 benzoquinone) concentration and flow carrier ph on the amperometric response of the g/cdh modified electrodes were investigated under flow conditions. from the calibration curves, recorded at two ph values (5.2 and ...201222809780
the translin-trax complex (c3po) is a ribonuclease in trna processing.conserved translin-trax complexes, also known as c3pos, have been implicated in many biological processes, but how they function remains unclear. recently, c3po was shown to be an endornase that promotes rna interference (rnai) in animal cells. here, we show that c3po does not play a significant role in rnai in the filamentous fungus neurospora crassa. instead, the neurospora c3po functions as an rnase that removes the 5' pre-trna fragments after the processing of pre-trnas by rnase p. in additi ...201222773104
cdr4 is the major contributor to azole resistance among four pdr5p-like abc transporters in neurospora crassa.pdr5p-like abc transporters play a significant role in azole resistance in saccharomyces cerevisiae and candida spp. most of filamentous fungal species have multiple pdr5p homologues. in this study, phylogenic analysis identified that filamentous fungi have at least two phylogenically distant groups of pdr5p homologues. one contains pmr1-like pdr5p homologues while the other contains both atrf-like and atrb-like pdr5p homologues. neurospora crassa has a total of four genes encoding pdr5p homolog ...201222749171
analysis of the sulfur-regulated control of the cystathionine γ-lyase gene of neurospora crassa.cystathionine γ-lyase plays a key role in the transsulfuration pathway through its primary reaction of catalyzing the formation of cysteine from cystathionine. the neurospora crassa cystathionine γ-lyase gene (cys-16(+)) is of particular interest in dissecting the regulation and dynamics of transsulfuration. the aim of this study was to determine the regulatory connection of cys-16(+) to the neurospora sulfur regulatory network. in addition, the cys-16(+) promoter was characterized with the goal ...201222748183
rna interference pathways in fungi: mechanisms and functions.rna interference (rnai) is a conserved eukaryotic gene regulatory mechanism that uses small noncoding rnas to mediate posttranscriptional/transcriptional gene silencing. the fission yeast schizosaccharomyces pombe and the filamentous fungus neurospora crassa have served as important model systems for rnai research. studies on these two organisms and other fungi have contributed significantly to our understanding of the mechanisms and functions of rnai in eukaryotes. in addition, surprisingly div ...201222746336
identification of genes required for alternative oxidase production in the neurospora crassa gene knockout library.the alternative oxidase (aox) of neurospora crassa transfers electrons from ubiquinol to oxygen. the enzyme is not expressed under normal conditions. however, when the function of the standard electron transport chain is compromised, aox is induced, providing cells with a means to continue respiration and growth. induction of the enzyme represents a form of retrograde regulation because aox is encoded by a nuclear gene that responds to signals produced from inefficiently functioning mitochondria ...201223173086
variation in mitochondrial genome primary sequence among whole-genome-sequenced strains of neurospora crassa.eighteen classical mutant strains of neurospora crassa were subject to whole genome sequence analysis and the mitochondrial genome is analyzed. overall, the mitochondrial genomes of the classical mutant strains are 99.45 to 99.98 % identical to the reference genome. two-thirds of the snps and three-fourths of indels identified in this analysis are shared among more than one strain. most of the limited variability in mitochondrial genome sequence is neutral with regard to protein structure. despi ...201223155504
a mouse neurodegenerative dynein heavy chain mutation alters dynein motility and localization in neurospora crassa.cytoplasmic dynein is responsible for the transport and delivery of cargoes in organisms ranging from humans to fungi. dysfunction of dynein motor machinery due to mutations in dynein or its activating complex dynactin can result in one of several neurological diseases in mammals. the mouse legs at odd angles (loa) mutation in the tail domain of the dynein heavy chain has been shown to lead to progressive neurodegeneration in mice. the mechanism by which the loa mutation affects dynein function ...201222991199
the pmr gene, encoding a ca2+-atpase, is required for calcium and manganese homeostasis and normal development of hyphae and conidia in neurospora crassa.the pmr gene is predicted to encode a ca(2+)-atpase in the secretory pathway. we examined two strains of neurospora crassa that lacked pmr: the δpmr strain, in which pmr was completely deleted, and pmr(rip), in which the gene was extensively mutated. both strains had identical, complex phenotypes. compared to the wild type, these strains required high concentrations of calcium or manganese for optimal growth and had highly branched, slow-growing hyphae. they conidiated poorly, and the shape and ...201222983986
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