scholarly journals Sulfonamide Inhibition Studies of the β-Class Carbonic Anhydrase CAS3 from the Filamentous Ascomycete Sordaria macrospora

Molecules ◽  
2020 ◽  
Vol 25 (5) ◽  
pp. 1036 ◽  
Author(s):  
Daniela Vullo ◽  
Ronny Lehneck ◽  
William A. Donald ◽  
Stefanie Pöggeler ◽  
Claudiu T. Supuran
Keyword(s):  
Carbonic Anhydrase ◽  
Model Organism ◽  
The Other ◽  
Antifungal Compounds ◽  
Inhibition Study ◽  
Sordaria Macrospora ◽  
Filamentous Ascomycete ◽  
Body Development ◽  
Fruiting Body Development ◽  
Inhibition Studies

A new β-class carbonic anhydrase was cloned and purified from the filamentous ascomycete Sordaria macrospora, CAS3. This enzyme has a higher catalytic activity compared to the other two such enzymes from this fungus, CAS1 and CAS2, which were reported earlier, with the following kinetic parameters: kcat of (7.9 ± 0.2) × 105 s−1, and kcat/Km of (9.5 ± 0.12) × 107 M−1∙s−1. An inhibition study with a panel of sulfonamides and one sulfamate was also performed. The most effective CAS3 inhibitors were benzolamide, brinzolamide, dichlorophnamide, methazolamide, acetazolamide, ethoxzolamide, sulfanilamide, methanilamide, and benzene-1,3-disulfonamide, with KIs in the range of 54–95 nM. CAS3 generally shows a higher affinity for this class of inhibitors compared to CAS1 and CAS2. As S. macrospora is a model organism for the study of fruiting body development in fungi, these data may be useful for developing antifungal compounds based on CA inhibition.

scholarly journals The Glyoxysomal Protease LON2 Is Involved in Fruiting-Body Development, Ascosporogenesis and Stress Resistance in Sordaria macrospora

2021 ◽  
Vol 7 (2) ◽  
pp. 82
Author(s):  
Antonia Werner ◽  
Kolja Otte ◽  
Gertrud Stahlhut ◽  
Leon M. Hanke ◽  
Stefanie Pöggeler
Keyword(s):  
Quality Control ◽  
Fruiting Body ◽  
Protein Quality ◽  
Model Organism ◽  
Protein Quality Control ◽  
Stress Conditions ◽  
Dual Function ◽  
Sordaria Macrospora ◽  
Body Development ◽  
Fruiting Body Development

Microbodies, including peroxisomes, glyoxysomes and Woronin bodies, are ubiquitous dynamic organelles that play important roles in fungal development. The ATP-dependent chaperone and protease family Lon that maintain protein quality control within the organelle significantly regulate the functionality of microbodies. The filamentous ascomycete Sordaria macrospora is a model organism for studying fruiting-body development. The genome of S. macrospora encodes one Lon protease with the C-terminal peroxisomal targeting signal (PTS1) serine-arginine-leucine (SRL) for import into microbodies. Here, we investigated the function of the protease SmLON2 in sexual development and during growth under stress conditions. Localization studies revealed a predominant localization of SmLON2 in glyoxysomes. This localization depends on PTS1, since a variant without the C-terminal SRL motif was localized in the cytoplasm. A ΔSmlon2 mutant displayed a massive production of aerial hyphae, and produced a reduced number of fruiting bodies and ascospores. In addition, the growth of the ΔSmlon2 mutant was completely blocked under mild oxidative stress conditions. Most of the defects could be complemented with both variants of SmLON2, with and without PTS1, suggesting a dual function of SmLON2, not only in microbody, but also in cytosolic protein quality control.

scholarly journals Anion Inhibition Studies of the β-Class Carbonic Anhydrase CAS3 from the Filamentous Ascomycete Sordaria macrospora

Metabolites ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 93
Author(s):  
Daniela Vullo ◽  
Ronny Lehneck ◽  
William A. Donald ◽  
Stefanie Pöggeler ◽  
Claudiu T. Supuran
Keyword(s):  
Catalytic Activity ◽  
Carbonic Anhydrase ◽  
Small Molecules ◽  
Small Molecule ◽  
Hydration Reaction ◽  
High Catalytic Activity ◽  
Sordaria Macrospora ◽  
Filamentous Ascomycete ◽  
Diethyl Dithiocarbamate ◽  
Inhibition Studies

CAS3 is a newly cloned cytosolic β-class carbonic anhydrase (CA, EC 4.2.1.1) from the filamentous ascomycete Sordaria macrospora. This enzyme has a high catalytic activity for the physiological CO2 hydration reaction and herein, we report the inhibition profile of CAS3 with anions and small molecules. The most effective CAS3 anions/small molecule inhibitors were diethyl-dithiocarbamate, sulfamide, sulfamate, phenyl boronic and phenyl arsonic acids, with KIs in the range of 0.89 mM–97 µM. Anions such as iodide, the pseudohalides, bicarbonate, carbonate, nitrate, nitrite, hydrogensulfide, stannate, selenate, tellurate, tetraborate, perrhenate, perruthenate, selenocyanide and trithiocarbonate were low millimolar CAS3 inhibitors. The light halides, sulfate, hydrogensulfite, peroxydisulfate, diphosphate, divanadate, perchlorate, tetrafluoroborate, fluorosulfonate and iminodisulfonate did not significantly inhibit this enzyme. These data may be useful for developing antifungals based on CA inhibition, considering the fact that many of the inhibitors reported here may be used as lead molecules and, by incorporating the appropriate organic scaffolds, potent nanomolar inhibitors could be developed.

scholarly journals Cloning the mating types of the heterothallic fungus Podospora anserina: developmental features of haploid transformants carrying both mating types.

Genetics ◽  
1991 ◽  
Vol 128 (3) ◽  
pp. 539-547 ◽  
Author(s):  
M Picard ◽  
R Debuchy ◽  
E Coppin
Keyword(s):  
Mating Type ◽  
Gene Replacement ◽  
Podospora Anserina ◽  
The Other ◽  
Mating Types ◽  
Body Development ◽  
Fruiting Body Development ◽  
Mat Locus ◽  
Functional Analyses ◽  
Developmental Features

Abstract DNAs that encode the mating-type functions (mat+ and mat-) of the filamentous fungus Podospora anserina were cloned with the use of the mating-type A probe from Neurospora crassa. Cloning the full mat information was ascertained through gene replacement experiments. Molecular and functional analyses of haploid transformants carrying both mating types lead to several striking conclusions. Mat+ mat- strains are dual maters. However, the resident mat information is dominant to the mat information added by transformation with respect to fruiting body development and ascus production. Moreover, when dual mating mat+ mat- strains are crossed to mat+ or mat- testers, there is strong selection, after fertilization, that leads to the loss from the mat+ mat- nucleus of the mat information that matches that of the tester. Finally, the mat locus contains at least two domains, one sufficient for fertilization, the other necessary for sporulation.

scholarly journals Functional Characterization of MAT1-1-Specific Mating-Type Genes in the Homothallic Ascomycete Sordaria macrospora Provides New Insights into Essential and Nonessential Sexual Regulators

2010 ◽  
Vol 9 (6) ◽  
pp. 894-905 ◽  
Author(s):  
V. Klix ◽  
M. Nowrousian ◽  
C. Ringelberg ◽  
J. J. Loros ◽  
J. C. Dunlap ◽  
...  
Keyword(s):  
Mating Type ◽  
Sexual Development ◽  
Fruiting Body ◽  
Content Type ◽  
Sordaria Macrospora ◽  
Body Development ◽  
Protein Encoding ◽  
Fruiting Body Development ◽  
Mating Type Genes ◽  
Domain Protein

ABSTRACT Mating-type genes in fungi encode regulators of mating and sexual development. Heterothallic ascomycete species require different sets of mating-type genes to control nonself-recognition and mating of compatible partners of different mating types. Homothallic (self-fertile) species also carry mating-type genes in their genome that are essential for sexual development. To analyze the molecular basis of homothallism and the role of mating-type genes during fruiting-body development, we deleted each of the three genes, SmtA-1 (MAT1-1-1), SmtA-2 (MAT1-1-2), and SmtA-3 (MAT1-1-3), contained in the MAT1-1 part of the mating-type locus of the homothallic ascomycete species Sordaria macrospora. Phenotypic analysis of deletion mutants revealed that the PPF domain protein-encoding gene SmtA-2 is essential for sexual reproduction, whereas the α domain protein-encoding genes SmtA-1 and SmtA-3 play no role in fruiting-body development. By means of cross-species microarray analysis using Neurospora crassa oligonucleotide microarrays hybridized with S. macrospora targets and quantitative real-time PCR, we identified genes expressed under the control of SmtA-1 and SmtA-2. Both genes are involved in the regulation of gene expression, including that of pheromone genes.

scholarly journals The pro1+ Gene From Sordaria macrospora Encodes a C6 Zinc Finger Transcription Factor Required for Fruiting Body Development

Genetics ◽  
1999 ◽  
Vol 152 (1) ◽  
pp. 191-199 ◽  
Author(s):  
Sandra Masloff ◽  
Stefanie Pöggeler ◽  
Ulrich Kück
Keyword(s):  
Zinc Finger ◽  
Fruiting Body ◽  
Molecular Mechanisms ◽  
Developmental Process ◽  
Open Reading Frame ◽  
Cosmid Library ◽  
Reading Frame ◽  
Sordaria Macrospora ◽  
Body Development ◽  
Fruiting Body Development

Abstract During sexual morphogenesis, the filamentous ascomycete Sordaria macrospora differentiates into multicellular fruiting bodies called perithecia. Previously it has been shown that this developmental process is under polygenic control. To further understand the molecular mechanisms involved in fruiting body formation, we generated the protoperithecia forming mutant pro1, in which the normal development of protoperithecia into perithecia has been disrupted. We succeeded in isolating a cosmid clone from an indexed cosmid library, which was able to complement the pro1- mutation. Deletion analysis, followed by DNA sequencing, subsequently demonstrated that fertility was restored to the pro1 mutant by an open reading frame encoding a 689-amino-acid polypeptide, which we named PRO1. A region from this polypeptide shares significant homology with the DNA-binding domains found in fungal C6 zinc finger transcription factors, such as the GAL4 protein from yeast. However, other typical regions of C6 zinc finger proteins, such as dimerization elements, are absent in PRO1. The involvement of the pro1+ gene in fruiting body development was further confirmed by trying to complement the mutant phenotype with in vitro mutagenized and truncated versions of the pro1 open reading frame. Southern hybridization experiments also indicated that pro1+ homologues are present in other sexually propagating filamentous ascomycetes.

New insights from an old mutant: SPADIX4 governs fruiting body development but not hyphal fusion in Sordaria macrospora

2016 ◽  
Vol 292 (1) ◽  
pp. 93-104 ◽  
Author(s):  
Ines Teichert ◽  
Miriam Lutomski ◽  
Ramona Märker ◽  
Minou Nowrousian ◽  
Ulrich Kück
Keyword(s):  
Fruiting Body ◽  
Hyphal Fusion ◽  
Sordaria Macrospora ◽  
Body Development ◽  
Fruiting Body Development
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