Msi1-like (MSIL) proteins, which are eukaryote-specific and contain a series of WD40 repeats, have pleiotropic roles in chromatin assembly, DNA damage repair, and regulation of nutrient/stress-sensing signaling pathways. In the fungal kingdom, the functions of MSIL proteins have been studied most intensively in the budding yeast model Saccharomyces cerevisiae, an ascomycete. Yet their functions are largely unknown in other fungi. Recently, an MSIL protein, Msl1, was discovered and functionally characterized in the pathogenic yeast Cryptococcus neoformans, a basidiomycete. Interestingly, MSIL proteins appear to have redundant and unique roles in both fungi, suggesting that MSIL proteins may have evolutionarily divergent roles in different parts of the fungal kingdom. In this review, we will describe the current findings regarding the role of MSIL proteins in fungi and discuss future directions for research on this topic.
Ascomycota ; Basidiomycota ; Chromatin Assembly and Disassembly ; Cryptococcus neoformans ; DNA Damage ; Fungi ; Histones ; Proteins ; Retinoblastoma ; Saccharomyces cerevisiae ; Saccharomycetales ; Yeasts

OBJECTIVETo identify the function of the gene encoding Neurospora crassa EAA33149.1 protein which has 46.85% similarity with Aspergillus niger phA gene.

METHODSThe bioinformatics analysis was conducted using the prediction algorithms SignalP v3.0, arginine and lysine propeptide cleavage sites in eukaryotic protein sequence prediction algorithms ProP 1.0 server, transmembrane domain prediction algorithms Tmpred and TMHMM v2.0, potential GP I-anchor sites prediction algorithm big-P I Predictor and the subcellular protein location prediction algorithms TargetP v1.01. According to the analysis results, the gene was cloned into Saccharomyces cerevisiae.

RESULTSThe signal peptide, the cleavage site and the secretion pathway were determined, and the expressed recombinant protein with 54,000 displayed phytase activity.

CONCLUSIONThe gene has been identified to encode N. crassa phyA.

Algorithms ; Amino Acid Sequence ; Computational Biology ; methods ; Fungal Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Neurospora crassa ; genetics ; Phytochrome A ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism

Kluyveromyces fragilis KCTC 7260 and Saccharomyces cerevisiae KCTC 7904, which both grew well in pear marc extract, were selected and their growth profiles and physiological functionalities were determined. Both of the selected yeasts established maximal growth by 20 hr of cultivation at 30degrees C in pear marc extract. The cell-free extracts showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 68.9% and 52.1%, respectively. The extracts also displayed 9.2 U/mL and 12.0 U/mL of protease activity, respectively.
Kluyveromyces ; Peptidyl-Dipeptidase A ; Pyrus ; Saccharomyces cerevisiae ; Yeasts

A continuous ethanol fermentation system composed of four-stage tank fermentors in series and with a total working volume of 4000 mL was established. The first fermentor was designated as the seed fermentor and the others for ethanol fermentation. A self-flocculating yeast strain developed by protoplast fusion of Saccharomyces cerevisiae and Schizosaccharomyces pombe was applied. Two-stage corn powder enzymatic hydrolyzate containing reducing sugar 100 g/L, together with 2.0 g/L (NH4)2HPO4 and KH2PO4, was used as yeast seed culture medium and fed into the seed fermentor at the dilution rate of 0.017h (-1). Meanwhile, the hydrolyzate containing reducing sugar 220 g/L, added with 1.5 g/L (NH4)2HPO4 and 2.5 g/L KH2PO4, was used as ethanol fermentation substrate and fed into the second fermentor at the dilution rates of 0.017, 0.025, 0.033, 0.040 and 0.050 h(-1) (based on the total working volume of the three fermentors), respectively. The chemostat states on which all of the monitoring parameters, including residual sugar, ethanol and yeast cell biomass concentrations, were maintained relatively constant were observed for seed cultivation and ethanol fermentations when the fermentation system was operated at the dilution rates of 0.017, 0.025, 0.033 and 0.050 h(-1). Yeast cells were observed being partly immobilized because significant yeast cell biomass concentration differences between the broth out of and inside the fermentors were detected. Moreover, the oscillations of residual sugar, ethanol and yeast cell biomass concentrations were observed when the fermentation system was operated at the dilution rate of 0.040 h(-1). The broth containing more than 12% (V/V) ethanol and less than 0.11% (W/V) residual reducing sugar and 0.35% (W/V) residual total sugar was produced when the dilution rate was controlled at no more than 0.033 h(-1). The ethanol productivity was calculated to be 3.32(g x L(-1) x h(-1)) for the dilution rate of 0.033 h(-1), which increased nearly 100% compared with that for conventional ethanol fermentation technologies using freely suspended yeast cells.
Bioreactors ; microbiology ; Carbohydrates ; Ethanol ; metabolism ; Fermentation ; Flocculation ; Saccharomyces cerevisiae ; metabolism ; Schizosaccharomyces ; metabolism ; Zea mays ; metabolism

A fermentation system composed of four airlift suspended-bed bioreactors in series and with a total working volume of 4800 mL was established. Continuous ethanol fermentation using self-flocculating yeast SPSC01, a fusant from Saccharomyces cerevisiae and Schizosaccharomyces pombe, and two-stage enzymatic hydrolyte of dry milling corn powder, was continuously run for 120 days. All of the backset distillage collected after distilling the final beer was used to mix the corn powder and no any other wastes except the solid residue of corn powder was discharged from the fermentation system, which guaranteed the distillage to be recycled at its maximum. The experimental results revealed that both ethanol and residual sugar in the final beer could be maintained relatively stable with their average levels of 93.6 and 7.9 g/L, respectively when the fermentation system was operated at the dilution rate of 0.05 h(-1). Parameter oscillations reported previously were also observed for the first and second bioreactors, but were effectively attenuated thereafter, which indicated that high yeast cell concentrations resulted from the self-immobilization of this special self-flocculating strain contributed to damp these oscillations. The monitoring of residual nitrogen and phosphor indicated that the accumulations of these nutritional elements occurred and the amount of these inorganic salts supplemented in the substrate should be decreased properly.
Bioreactors ; microbiology ; Ethanol ; metabolism ; Fermentation ; Flocculation ; Immobilization ; Saccharomyces cerevisiae ; metabolism ; Schizosaccharomyces ; metabolism ; Zea mays ; metabolism

Peanut yield and quality are seriously affected by pod rot pathogens worldwide, especially in China in recent years. The goals of this study are to analyze the structure of fungal communities of peanut pod rot in soil in three peanut cultivars and the correlation of pod rot with environmental variables using 454 pyrosequencing. A total of 46,723 internal transcribed spacer high-quality sequences were obtained and grouped into 1,706 operational taxonomic units at the 97% similarity cut-off level. The coverage, rank abundance, and the Chao 1 and Shannon diversity indices of the operational taxonomic units were analyzed. Members of the phylum Ascomycota were dominant, such as Fusarium, Chaetomium, Alternaria, and Sordariomycetes, followed by Basidiomycota. The results of the heatmap and redundancy analysis revealed significant variation in the composition of the fungal community among the three cultivar samples. The environmental conditions in different peanut cultivars may also influence on the structure of the fungal community. The results of this study suggest that the causal agent of peanut pod rot may be more complex, and cultivars and environmental conditions are both important contributors to the community structure of peanut pod rot fungi.
Alternaria ; Arachis* ; Ascomycota ; Basidiomycota ; Chaetomium ; China* ; Fungi* ; Fusarium ; Soil*

The flocculating yeast strain SPSC01 is a fusant strain of Saccharomyces cerevisiae and Schizosaccharomyces pombe. The use of SPSC01 to absorb Cr(VI) from Cr(VI) containing aqueous solution would greatly reduce the cost of post-adsorption separation, since the superior flocculating property of SPSC01 would allow easy separation of the Cr(VI)-biomass from the solution. In order to investigate the effects of flocculating proteins on Cr(VI) reduction and absorption by SPSC01, the absorption behaviors of SPSC01 and its parental strains were compared. The results showed that Cr(VI) removal rate of SPSC01 was almost the same as that of S. pombe, which also has flocculating ability, but was faster than that of S. cerevisiae, which has no flocculating ability. When the system reached equilibrium, the amount of total Cr adsorbed by S. pombe, SPSC01 and S. cerevisiae were 68.8%, 48.6% and 37.5%, respectively. This showed that flocculation was beneficial to Cr(VI) reduction and adsorption, and suggested that focculating proteins may play a role in enhancing the Cr(VI) adsorption capacity of SPSC01 and S. pombe. We investigated the mechanism of Cr(VI) adsorption by SPSC01 using chemical modification and FTIR. The results indicated that the major functional groups (amino, carboxyl and amide) of surface proteins may contribute to the absorption of Cr(VI).
Adsorption ; Biodegradation, Environmental ; Chromium ; isolation & purification ; Flocculation ; Saccharomyces cerevisiae ; metabolism ; Schizosaccharomyces ; metabolism ; Surface Properties ; Water Pollutants, Chemical ; isolation & purification

Circadian clocks are the internal time-keeping mechanisms for organisms to synchronize their cellular and physiological processes to the daily light/dark cycles. The molecular mechanisms underlying circadian clocks are remarkably similar in eukaryotes. Neurospora crassa, a filamentous fungus, is one of the best understood model organisms for circadian research. In recent years, accumulating data have revealed complex regulation in the Neurospora circadian clock at transcriptional, posttranscriptional, post-translational and epigenetic levels. Here we review the recent progress towards our understanding of the molecular mechanism of the Neurospora circadian oscillator. These advances have provided novel insights and furthered our understanding of the mechanism of eukaryotic circadian clocks.
Circadian Clocks ; Epigenomics ; Neurospora ; genetics ; metabolism ; physiology ; Neurospora crassa ; genetics ; metabolism ; physiology

A nucleoside diphosphate-linked moiety X (Nudix) hydrolase-like gene, YSA1, has been identified as one of the gromwell plant extract-responsive genes in Cryptococcus neoformans. Ysa1 is known to control intracellular concentrations of ADP-ribose or O-acetyl-ADP-ribose, and has diverse biological functions, including the response to oxidative stress in the ascomycete yeast, Saccharomyces cerevisiae. In this study, we characterized the role of YSA1 in the stress response and adaptation of the basidiomycete yeast, C. neoformans. We constructed three independent deletion mutants for YSA1, and analyzed their mutant phenotypes. We found that ysa1 mutants did not show increased sensitivity to reactive oxygen species-producing oxidative damage agents, such as hydrogen peroxide and menadione, but exhibited increased sensitivity to diamide, which is a thiol-specific oxidant. Ysa1 was dispensable for the response to most environmental stresses, such as genotoxic, osmotic, and endoplasmic reticulum stress. In conclusion, modulation of YSA1 may regulate the cellular response and adaptation of C. neoformans to certain oxidative stresses and contribute to the evolution of antifungal drug resistance.
Adenosine Diphosphate Ribose ; Ascomycota ; Basidiomycota ; Cryptococcus neoformans* ; Cryptococcus* ; Diamide ; Drug Resistance, Fungal ; Endoplasmic Reticulum Stress ; Hydrogen Peroxide ; Lithospermum ; O-Acetyl-ADP-Ribose ; Oxidative Stress* ; Oxygen ; Phenotype ; Plants ; Saccharomyces cerevisiae ; Vitamin K 3 ; Yeasts

Ca2+ at 1.64 mmol/L markedly increased ethanol tolerance of a self-flocculating fusant of Schizosaccharomyces pombe and Saccharomyces cerevisiae. After 9 h of exposure to 20% (V/V) ethanol at 30 degrees C , no viability remained for the control whereas 50.0% remained for the cells both grown and incubated with ethanol in Ca2+ -added medium. Furthermore, when subjected to 15% (V/V) ethanol at 30 degrees C, the equilibrium nucleotide concentration and plasma membrane permeability coefficient (P' ) of the cells both grown and incubated with ethanol in Ca2+ -added medium accounted for only 50.0% and 29.3% those of the control respectively, indicating that adding Ca2+ can markedly reduce plasma membrane permeability of yeast cells under ethanol stress as compared with the control. Meanwhile, high viability levels acquired by the addition of Ca2+ exactly corresponded to the striking decreases in extracellular nucleotide concentration and P' achieved with identical approach. Therefore, the enhancing effect of Ca2+ on ethanol tolerance of this strain is closely related to its ability to decrease plasma membrane permeability of yeast cells subjected to ethanol stress.
Calcium ; pharmacology ; Cell Membrane ; drug effects ; metabolism ; Cell Membrane Permeability ; drug effects ; Ethanol ; pharmacology ; Saccharomyces cerevisiae ; drug effects ; growth & development ; metabolism ; Schizosaccharomyces ; drug effects ; growth & development ; metabolism ; Temperature