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Aspergillus nidulans

Luiza Cesca Piva, Janice Lisboa De Marco, Lidia Maria Pepe de Moraes, Viviane Castelo Branco Reis, Fernando Araripe Gonçalves Torres
We have investigated the use of the gene coding for acetamidase (amdS) as a recyclable dominant marker for the methylotrophic yeast Komagataella phaffii in order to broaden its genetic toolbox. First, the endogenous constitutive AMD2 gene (a putative acetamidase) was deleted generating strain LA1. A cassette (amdSloxP) was constructed bearing a codon-optimized version of the Aspergillus nidulans amdS gene flanked by loxP sites for marker excision with Cre recombinase. This cassette was successfully tested as a dominant selection marker for transformation of the LA1 strain after selection on plates containing acetamide as a sole nitrogen source...
February 12, 2018: Applied Microbiology and Biotechnology
I Al Salmi, A M Metry, F Al Ismaili, A Hola, M Al Riyami, F Khamis, S Al-Abri
BACKGROUND: Deceased and live-related renal transplants (RTXs) are widespread approved methods throughout the world. In addition, commercial RTX has become popular in certain places driven by financial greed. METHOD: This is a retrospective, descriptive study was done at the Royal Hospital (RH) from 2013 to 2015. The data have been collected from the national Oman kidney transplant registry. All transplant cases retrieved were divided into two groups, Live-related-RTX performed in Oman and commercial-unrelated-RTX performed abroad...
February 8, 2018: International Journal of Infectious Diseases: IJID
Ashraf A Kadry, Amira M El-Ganiny, Rasha A Mosbah, Susan G W Kaminskyj
Systemic human fungal infections are increasingly common. Aspergillus species cause most of the airborne fungal infections. Life-threatening invasive aspergillosis was formerly found only in immune-suppressed patients, but recently some strains of A. fumigatus have become primary pathogens. Many fungal cell wall components are absent from mammalian systems, so they are potential drug targets. Cell-wall-targeting drugs such as echinocandins are used clinically, although echinocandin-resistant strains were discovered shortly after their introduction...
October 9, 2017: Medical Mycology: Official Publication of the International Society for Human and Animal Mycology
Jasmine Pham, Remco Stam, Victor Martinez Heredia, Michael Csukai, Edgar Huitema
Phytophthora spp. cause devastating disease epidemics on important crop plants and pose a grave threat to global crop production. Critically, Phytophthora pathogens represent a distinct evolutionary lineage in which pathogenicity has been acquired independently. Therefore, there is an urgent need to understand and disrupt the processes that drive infection, if we aspire to defeat oomycete parasites in the field. One area that has received little attention thus far in this respect is the regulation of Phytophthora gene expression during infection...
February 8, 2018: Molecular Plant-microbe Interactions: MPMI
Yibo Hu, Yuqi Qin, Guodong Liu
Transcriptional regulation has important roles in various biological processes (e.g., development and metabolism) in filamentous fungi. However, regulatory interactions between transcription factors (TFs) and their target genes in these species have only been described in different forms by primary scientific literature, which limits the integrated analysis of these data. Here, we extensively curated the reported transcriptional regulatory interactions in Aspergillus nidulans and Neurospora crassa. For each interaction, the identifiers of involved proteins or genes were unified, and the types of supporting experiments were recorded...
2018: Frontiers in Microbiology
Miriam Kolog Gulko, Gabriele Heinrich, Carina Gross, Blagovesta Popova, Oliver Valerius, Piotr Neumann, Ralf Ficner, Gerhard H Braus
The transition from vegetative growth to multicellular development represents an evolutionary hallmark linked to an oxidative stress signal and controlled protein degradation. We identified the Sem1 proteasome subunit, which connects stress response and cellular differentiation. The sem1 gene encodes the fungal counterpart of the human Sem1 proteasome lid subunit and is essential for fungal cell differentiation and development. A sem1 deletion strain of the filamentous fungus Aspergillus nidulans is able to grow vegetatively and expresses an elevated degree of 20S proteasomes with multiplied ATP-independent catalytic activity compared to wildtype...
February 5, 2018: PLoS Genetics
Lu Zhou, Minoas Evangelinos, Valentin Wernet, Antonia F Eckert, Yuji Ishitsuka, Reinhard Fischer, G Ulrich Nienhaus, Norio Takeshita
Polarized growth of filamentous fungi requires continuous transport of biomolecules to the hyphal tip. To this end, construction materials are packaged in vesicles and transported by motor proteins along microtubules and actin filaments. We have studied these processes with quantitative superresolution localization microscopy of live Aspergillus nidulans cells expressing the photoconvertible protein mEosFPthermo fused to the chitin synthase ChsB. ChsB is mainly located at the Spitzenkörper near the hyphal tip and produces chitin, a key component of the cell wall...
January 2018: Science Advances
Candace R Alexander, Douglas W Dingman, Neil P Schultes, George S Mourad
Two nucleobase transporters encoded in the genome of the Honey bee bacterial pathogen Paenibacillus larvae belong to the azaguanine-like transporters and are referred to as PlAzg1 and PlAzg2. PlAzg1 & 2 display significant amino acid sequence similarity, share predicted secondary structures and functional sequence motifs with two Escherichia coli nucleobase cation symporter 2 (NCS2) members; adenine permease (EcAdeP) and guanine-hypoxanthine permease EcGhxP. However, similarity does not define function. Heterologous complementation and functional analysis using nucleobase transporter deficient Saccharomyces cerevisiae strains revealed that PlAzg1 transports, adenine, hypoxanthine, xanthine and uracil, while PlAzg2 transports adenine, guanine, hypoxanthine, xanthine, cytosine and uracil...
January 29, 2018: FEMS Microbiology Letters
Ebru Alazi, Tim Knetsch, Marcos Di Falco, Ian D Reid, Mark Arentshorst, Jaap Visser, Adrian Tsang, Arthur F J Ram
The transcription factor GaaR is needed for the expression of genes required for pectin degradation and transport and catabolism of the main degradation product, D-galacturonic acid (GA) in Aspergillus niger. In this study, we used the strong constitutive gpdA promoter of Aspergillus nidulans to overexpress gaaR in A. niger. Overexpression of gaaR resulted in an increased transcription of the genes encoding pectinases, (putative) GA transporters, and catabolic pathway enzymes even under non-inducing conditions, i...
January 24, 2018: Applied Microbiology and Biotechnology
S K Shanuja, S Iswarya, A Gnanamani
The present study explores UVB protective role of a melanin precursor namely DHICA (5,6- Dihydroxyindole-2-carboxylic acid) expressed by the marine imperfect fungus Aspergillus nidulans. In brief, A. nidulans grown in a modified growth medium for the period of 5 days at 25 °C under shaking conditions and the extracellular medium free from fungal biomass used for the extraction of DHICA. The extracted DHICA further exposed to partial purification and subjected to UVB protection studies using HaCaT cells and Balb/c mice independently...
January 19, 2018: Journal of Photochemistry and Photobiology. B, Biology
Yang Xia, Haijun Yu, Zhemin Zhou, Naoki Takaya, Shengmin Zhou, Ping Wang
Most eukaryotic peroxiredoxins (Prxs) are readily inactivated by high concentration of hydrogen peroxide (H₂O₂) during catalysis due to their "GGLG" and "YF" motif. However, such oxidative stress sensitive motifs were not found in the previously identified filamentous fungal Prxs. Additionally, the information of filamentous fungal Prxs is limited and fragmentary. Herein, we cloned and gained insight into Aspergillus nidulans Prx, (An.PrxA), in the aspects of protein properties, catalysis characteristics and especially in H₂O₂ tolerability...
January 24, 2018: Journal of Microbiology and Biotechnology
Liangkun Long, Haoyuan Zhao, Dafan Ding, Meijuan Xu, Shaojun Ding
Feruloyl esterase (FAE)-encoding genes AnfaeA and AnfaeB were isolated from Aspergillus niger 0913. For overexpression of the two genes in Trichoderma reesei, constitutive and inductive expression plasmids were constructed based on parental plasmid pAg1-H3. The constructed plasmids contained AnfaeA or AnfaeB gene under the control of glyceraldehyde-3-phosphate dehydrogenase A gene (gpdA) promoter (from A. nidulans) or cellobiohydrolases I (cbh I) gene promoter (from T. reesei), and cbh I terminator from T. reesei...
January 18, 2018: Bioprocess and Biosystems Engineering
Raul Machado, André da Costa, Dina M Silva, Andreia C Gomes, Margarida Casal, Vitor Sencadas
Antimicrobial materials have become relevant for local therapies preventing microbial resistance induced by systemic antibiotic treatments. This work reports the development of electrospun poly(lactic acid) (PLLA) nanofiber membranes loaded with bovine lactoferrin (bLF) up to 20 wt%. The membranes present smooth and nondefective fibers with mean diameters between 717 ± 197 and 495 ± 127 nm, and an overall porosity of ≈80%. The hydrophobicity of the PLLA membranes is reduced by the presence of bLF. The release profile of bLF correlates with an anomalous transport model, with 17...
January 15, 2018: Macromolecular Bioscience
Christina S Nødvig, Jakob B Hoof, Martin E Kogle, Zofia D Jarczynska, Jan Lehmbeck, Dorte K Klitgaard, Uffe H Mortensen
CRISPR-Cas9 technologies are revolutionizing fungal gene editing. Here we show that survival of specific Cas9/sgRNA mediated DNA double strand breaks (DSBs) depends on the non-homologous end-joining, NHEJ, DNA repair pathway and we use this observation to develop a tool to assess protospacer efficiency in Aspergillus nidulans. Moreover, we show that in NHEJ deficient strains, highly efficient marker-free gene targeting can be performed. Indeed, we show that even single-stranded oligo nucleotides efficiently works as repair templates of specific Cas9/sgRNA induced DNA DSBs in A...
January 8, 2018: Fungal Genetics and Biology: FG & B
Yu Nakashima, Takahiro Mori, Hitomi Nakamura, Takayoshi Awakawa, Shotaro Hoshino, Miki Senda, Toshiya Senda, Ikuro Abe
Non-heme iron and α-ketoglutarate (αKG) oxygenases catalyze remarkably diverse reactions using a single ferrous ion cofactor. A major challenge in studying this versatile family of enzymes is to understand their structure-function relationship. AusE from Aspergillus nidulans and PrhA from Penicillium brasilianum are two highly homologous Fe(II)/αKG oxygenases in fungal meroterpenoid biosynthetic pathways that use preaustinoid A1 as a common substrate to catalyze divergent rearrangement reactions to form the spiro-lactone in austinol and cycloheptadiene moiety in paraherquonin, respectively...
January 9, 2018: Nature Communications
Mariane Paludetti Zubieta, Fabiano Jares Contesini, Marcelo Ventura Rubio, Any Elisa de Souza Schmidt Gonçalves, Jaqueline Aline Gerhardt, Rolf Alexander Prade, André Ricardo de Lima Damasio
Filamentous fungi are robust cell factories and have been used for the production of large quantities of industrially relevant enzymes. However, the production levels of heterologous proteins still need to be improved. Therefore, this article aimed to investigate the global proteome profiling of Aspergillus nidulans recombinant strains in order to understand the bottlenecks of heterologous enzymes production. About 250, 441 and 424 intracellular proteins were identified in the control strain Anid_pEXPYR and in the recombinant strains Anid_AbfA and Anid_Cbhl respectively...
January 8, 2018: Microbial Biotechnology
James W Dohn, Alexander W Grubbs, C Elizabeth Oakley, Berl R Oakley
Technical advances in Aspergillus nidulans enable relatively easy deletion of genomic sequences, insertion of sequences into the genome and alteration of genomic sequences. To extend the power of this system we wished to create strains with several selectable markers in a common genetic background to facilitate multiple, sequential transformations. We have developed an approach, using the recycling of the pyrG selectable marker, that has allowed us to create new deletions of the biA, pabaA, choA, and lysB genes...
January 5, 2018: Fungal Genetics and Biology: FG & B
Andrew M Fry, Richard Bayliss, Joan Roig
Genetic studies in yeast and Drosophila led to identification of cyclin-dependent kinases (CDKs), Polo-like kinases (PLKs) and Aurora kinases as essential regulators of mitosis. These enzymes have since been found in the majority of eukaryotes and their cell cycle-related functions characterized in great detail. However, genetic studies in another fungal species, Aspergillus nidulans, identified a distinct family of protein kinases, the NEKs, that are also widely conserved and have key roles in the cell cycle, but which remain less well studied...
2017: Frontiers in Cell and Developmental Biology
Mark S Gresnigt, Katharina L Becker, Floris Leenders, M Fernanda Alonso, Xiaowen Wang, Jacques F Meis, Judith M Bain, Lars P Erwig, Frank L van de Veerdonk
Invasive aspergillosis mainly occurs in immunocompromised patients and is commonly caused by Aspergillus fumigatus, while A.nidulans is rarely the causative agent. However, in chronic granulomatous disease (CGD) patients, A. nidulans is a frequent cause of invasive aspergillosis and is associated with higher mortality. Immune recognition of A. nidulans was compared to A. fumigatus to offer an insight into why A. nidulans infections are prevalent in CGD. Live cell imaging with J774A.1 macrophage-like cells and LC3-GFP-mCherry bone marrow-derived macrophages (BMDMs) revealed that phagocytosis of A...
December 16, 2017: Journal of Innate Immunity
Jessica J Talbot, Paul Thompson, Larry Vogelnest, Vanessa R Barrs
Aspergillosis is a major cause of severe respiratory disease in birds. The prevalence of cryptic section Fumigati and other non-Aspergillus fumigatus species as causative agents is unknown. Species identity was determined in 30 isolates from affected birds from zoos, pet birds and poultry by PCR of the ITS1-5.8S-ITS2 and partial β-tubulin genes. The most prevalent isolate was A. fumigatus sens. str. in 87% (26) cases. Other Aspergillus species were identified in 13% (4) cases, including A. restrictus (1), A...
December 8, 2017: Medical Mycology: Official Publication of the International Society for Human and Animal Mycology
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