scholarly journals Asexual Development Is Increased in Neurospora crassa cat-3-Null Mutant Strains

2003 ◽  
Vol 2 (4) ◽  
pp. 798-808 ◽  
Author(s):  
Shaday Michán ◽  
Fernando Lledías ◽  
Wilhelm Hansberg

ABSTRACT We use asexual development of Neurospora crassa as a model system with which to determine the causes of cell differentiation. Air exposure of a mycelial mat induces hyphal adhesion, and adherent hyphae grow aerial hyphae that, in turn, form conidia. Previous work indicated the development of a hyperoxidant state at the start of these morphogenetic transitions and a large increase in catalase activity during conidiation. Catalase 3 (CAT-3) increases at the end of exponential growth and is induced by different stress conditions. Here we analyzed the effects of cat-3-null strains on growth and asexual development. The lack of CAT-3 was not compensated by other catalases, even under oxidative stress conditions, and cat-3RIP colonies were sensitive to H2O2, indicating that wild-type (Wt) resistance to external H2O2 was due to CAT-3. cat-3RIP colonies grown in the dark produced high levels of carotenes as a consequence of oxidative stress. Light exacerbated oxidative stress and further increased carotene synthesis. In the cat-3RIP mutant strain, increased aeration in liquid cultures led to increased hyphal adhesion and protein oxidation. Compared to the Wt, the cat-3RIP mutant strain produced six times more aerial hyphae and conidia in air-exposed mycelial mats, as a result of longer and more densely packed aerial hyphae. Protein oxidation in colonies was threefold higher and showed more aerial hyphae and conidia in mutant strains than did the Wt. Results indicate that oxidative stress due to lack of CAT-3 induces carotene synthesis, hyphal adhesion, and more aerial hyphae and conidia.

Microbiology ◽  
2006 ◽  
Vol 152 (6) ◽  
pp. 1687-1700 ◽  
Author(s):  
Jesús Delgado-Jarana ◽  
Sonia Sousa ◽  
Fran González ◽  
Manuel Rey ◽  
Antonio Llobell

Trichoderma harzianum is a widespread mycoparasitic fungus, able to successfully colonize a wide range of substrates under different environmental conditions. Transcript profiling revealed a subset of genes induced in T. harzianum under hyperosmotic shock. The hog1 gene, a homologue of the MAPK HOG1 gene that controls the hyperosmotic stress response in Saccharomyces cerevisiae, was characterized. T. harzianum hog1 complemented the hog1Δ mutation in S. cerevisiae, but showed different features to yeast alleles: improved osmoresistance by expression of the hog1 allele and a lack of lethality when the hog1 F315S allele was overexpressed. ThHog1 protein was phosphorylated in T. harzianum under different stress conditions such as hyperosmotic or oxidative stress, among others. By using a ThHog1-GFP fusion, the protein was shown to be localized in nuclei under these stress conditions. Two mutant strains of T. harzianum were constructed: one carrying the hog1 F315S allele, and a knockdown hog1-silenced strain. The silenced strain was highly sensitive to osmotic stress, and showed intermediate levels of resistance against oxidative stress, indicating that the main role of ThHog1 protein is in the hyperosmotic stress response. Stress cross-resistance experiments showed evidences of a secondary role of ThHog1 in oxidative stress. The strain carrying the hog1 F315S allele was highly resistant to the calcineurin inhibitor cyclosporin A, which suggests the existence of links between the two pathways. The two mutant strains showed a strongly reduced antagonistic activity against the plant pathogens Phoma betae and Colletotrichum acutatum, which points to a role of ThHog1 protein in fungus–fungus interactions.


2006 ◽  
Vol 188 (14) ◽  
pp. 5325-5330 ◽  
Author(s):  
Praveen Alamuri ◽  
Nalini Mehta ◽  
Andrew Burk ◽  
Robert J. Maier

ABSTRACT Transcription of both chromosomal and extrachromosomally introduced nifS was regulated (up-expressed) by oxygen or by supplemental iron conditions. This up-expression was not observed in a fur mutant strain background or when an iron chelator was added. Iron-bound Fur (but not apo-Fur) recognized the nifS promoter, and Fur bound significantly farther upstream (−155 bp to −190 bp and −210 to −240 bp) in the promoter than documented Helicobacter pylori Fur binding regions. This binding was stronger than Fur recognition of the flgE or napA promoter and includes a Fur recognition sequence common to the H. pylori pfr and sodB upstream areas. Studies of Fur-regulated genes in H. pylori have indicated that apo-Fur acts as a repressor, but our results demonstrate that iron-bound Fur activates (nifS) transcription.


2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Kathrin Ulrich ◽  
Blanche Schwappach ◽  
Ursula Jakob

AbstractThiol-based redox switches evolved as efficient post-translational regulatory mechanisms that enable individual proteins to rapidly respond to sudden environmental changes. While some protein functions need to be switched off to save resources and avoid potentially error-prone processes, protective functions become essential and need to be switched on. In this review, we focus on thiol-based activation mechanisms of stress-sensing chaperones. Upon stress exposure, these chaperones convert into high affinity binding platforms for unfolding proteins and protect cells against the accumulation of potentially toxic protein aggregates. Their chaperone activity is independent of ATP, a feature that becomes especially important under oxidative stress conditions, where cellular ATP levels drop and canonical ATP-dependent chaperones no longer operate. Vice versa, reductive inactivation and substrate release require the restoration of ATP levels, which ensures refolding of client proteins by ATP-dependent foldases. We will give an overview over the different strategies that cells evolved to rapidly increase the pool of ATP-independent chaperones upon oxidative stress and provide mechanistic insights into how stress conditions are used to convert abundant cellular proteins into ATP-independent holding chaperones.


Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 823
Author(s):  
Cristiana Mateus ◽  
Ana Rita Nunes ◽  
Mónica Oleastro ◽  
Fernanda Domingues ◽  
Susana Ferreira

Aliarcobacter butzleri is an emergent enteropathogen that can be found in a range of environments. This bacterium presents a vast repertoire of efflux pumps, such as the ones belonging to the resistance nodulation cell division family, which may be associated with bacterial resistance, as well as virulence. Thus, this work aimed to evaluate the contribution of three RND efflux systems, AreABC, AreDEF and AreGHI, in the resistance and virulence of A. butzleri. Mutant strains were constructed by inactivation of the gene that encodes the inner membrane protein of these systems. The bacterial resistance profile of parental and mutant strains to several antimicrobials was assessed, as was the intracellular accumulation of the ethidium bromide dye. Regarding bacterial virulence, the role of these three efflux pumps on growth, strain fitness, motility, biofilm formation ability, survival in adverse conditions (oxidative stress and bile salts) and human serum and in vitro adhesion and invasion to Caco-2 cells was evaluated. We observed that the mutants from the three efflux pumps were more susceptible to several classes of antimicrobials than the parental strain and presented an increase in the accumulation of ethidium bromide, indicating a potential role of the efflux pumps in the extrusion of antimicrobials. The mutant strains had no bacterial growth defects; nonetheless, they presented a reduction in relative fitness. For the three mutants, an increase in the susceptibility to oxidative stress was observed, while only the mutant for AreGHI efflux pump showed a relevant role in bile stress survival. All the mutant strains showed an impairment in biofilm formation ability, were more susceptible to human serum and were less adherent to intestinal epithelial cells. Overall, the results support the contribution of the efflux pumps AreABC, AreDEF and AreGHI of A. butzleri to antimicrobial resistance, as well as to bacterial virulence.


Genetics ◽  
1999 ◽  
Vol 151 (1) ◽  
pp. 107-117
Author(s):  
Qi Yang ◽  
Katherine A Borkovich

Abstract Heterotrimeric G proteins, consisting of α, β, and γ subunits, transduce environmental signals through coupling to plasma membrane-localized receptors. We previously reported that the filamentous fungus Neurospora crassa possesses a Gα protein, GNA-1, that is a member of the Gαi superfamily. Deletion of gna-1 leads to defects in apical extension, differentiation of asexual spores, sensitivity to hyperosmotic media, and female fertility. In addition, Δgna-1 strains have lower intracellular cAMP levels under conditions that promote morphological abnormalities. To further define the function of GNA-1 in signal transduction in N. crassa, we examined properties of strains with mutationally activated gna-1 alleles (R178C or Q204L) as the only source of GNA-1 protein. These mutations are predicted to inhibit the GTPase activity of GNA-1 and lead to constitutive signaling. In the sexual cycle, gna-1R178C and gna-1Q204L strains are female-fertile, but produce fewer and larger perithecia than wild type. During asexual development, gna-1R178C and gna-1Q204L strains elaborate abundant, long aerial hyphae, produce less conidia, and possess lower levels of carotenoid pigments in comparison to wild-type controls. Furthermore, gna-1R178C and gna-1Q204L strains are more sensitive to heat shock and exposure to hydrogen peroxide than wild-type strains, while Δgna-1 mutants are more resistant. In contrast to Δgna-1 mutants, gna-1R178C and gna-1Q204L strains have higher steady-state levels of cAMP than wild type. The results suggest that GNA-1 possesses several Gβγ-independent functions in N. crassa. We propose that GNA-1 mediates signal transduction pathway(s) that regulate aerial hyphae development and sensitivity to heat and oxidative stresses, possibly through modulation of cAMP levels.


Nanoscale ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 3855-3863 ◽  
Author(s):  
Namrata Singh ◽  
Mohammed Azharuddin Savanur ◽  
Shubhi Srivastava ◽  
Patrick D'Silva ◽  
Govindasamy Mugesh

Multi-enzyme mimetic Mn3O4 nanoflowers (Mp) modulate the redox state of mammalian cells without altering the cellular antioxidant machinery under oxidative stress conditions.


2016 ◽  
Vol 12 (2) ◽  
pp. 391-403 ◽  
Author(s):  
Federico Lopez-Moya ◽  
David Kowbel ◽  
Ma José Nueda ◽  
Javier Palma-Guerrero ◽  
N. Louise Glass ◽  
...  

In this work we performed the first transcriptional study of a filamentous fungus (N. crassa) in response to chitosan.


1972 ◽  
Vol 19 (3) ◽  
pp. 191-204 ◽  
Author(s):  
J. Weijer ◽  
N. V. Vigfusson

SUMMARYMutations giving rise to sexual sterility were induced in Neurospora crassa macroconidia by ultraviolet-light irradiation. Thirty mutants were isolated on the basis of their male sterility in crosses with a wild-type strain. When used as the male parent these mutants exhibited a wide spectrum of sexual behaviour patterns ranging from the production of only small brown protoperithecia (complete male sterility) to the production of large and normally pigmented perithecia but with an undeveloped ostiole and very few if any spores. For many of the mutants the behaviour pattern is different when the strain is used as the female parent. Segregation data reveal that none of these mutants represent mutations of the mating-type locus. These findings suggest that the sexual development cycle is blocked at various stages in the different mutant strains. All attempts to restore fertility by supplying various additives to the medium or by varying the incubation time and temperature were unsuccessful. Conidial viability tests carried out on many of the strains revealed no abnormality in this respect. The aberrant segregation patterns exhibited by many of the mutants are discussed.


2010 ◽  
Vol 49 ◽  
pp. S134
Author(s):  
Fernando Toshio Ogata ◽  
Wagner Luiz Batista ◽  
Tarsis Gestira Ferreira ◽  
Roberto Jun Arai ◽  
Junji Yodoi ◽  
...  

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