Journal: Frontiers in Microbiology

Article Title: Differentially Regulated Transcription Factors and ABC Transporters in a Mitochondrial Dynamics Mutant Can Alter Azole Susceptibility of Aspergillus fumigatus

doi: 10.3389/fmicb.2020.01017

Figure Lengend Snippet: Analysis of differentially expressed genes in the Δ dnm1 mgm1 tetOn mutant. (A) The transcriptome of wild type (wt) and Δ dnm1 mgm1 tetOn were analyzed with RNA sequencing in triplicates. Differentially expressed genes were subsequently identified with Cuffdiff 32 . 621 genes were upregulated and 628 genes were downregulated ( p ≤ 0.007). (B,C) Significantly overrepresented (B) and underrepresented (C) GO terms (goslim_generic subset; p -value ≤ 0.05) for molecular function (black bars) and biological process (gray bars). The bars indicate the percentage of identified genes of all analyzed A. fumigatus genes associated with each GO slim term. The absolute number of identified genes is shown on top of each bar.

Article Snippet: For identification of the ABC and MFS transporters as well as the transcription factors which are up- and downregulated in Δ dnm1 mgm1 tetOn the differentially expressed genes were compared with the genes coding the indicated proteins using the free software Knime .

Techniques: Mutagenesis, RNA Sequencing

Journal: Frontiers in Microbiology

Article Title: Differentially Regulated Transcription Factors and ABC Transporters in a Mitochondrial Dynamics Mutant Can Alter Azole Susceptibility of Aspergillus fumigatus

doi: 10.3389/fmicb.2020.01017

Figure Lengend Snippet: Up- and downregulated genes Δ dnm1 mgm1 tetOn mutant with a presumed role in mitochondrial biology, transmembrane transport, stress response and ergosterol metabolism. The charts show the number of up- and downregulated genes that belong to the indicated GO terms: (A) mitochondrion (GO:0005739) and mitochondrion organization (GO:0007005); (B) transmembrane transport (GO:0055085) and transmembrane transporter activity (GO:0022857); (C) response to stress (GO:0006950); (D) ergosterol metabolic process (GO:0008204). The absolute number of genes associated with the indicated GO terms is shown in charts.

Article Snippet: For identification of the ABC and MFS transporters as well as the transcription factors which are up- and downregulated in Δ dnm1 mgm1 tetOn the differentially expressed genes were compared with the genes coding the indicated proteins using the free software Knime .

Techniques: Mutagenesis, Activity Assay

Journal: Frontiers in Microbiology

Article Title: Differentially Regulated Transcription Factors and ABC Transporters in a Mitochondrial Dynamics Mutant Can Alter Azole Susceptibility of Aspergillus fumigatus

doi: 10.3389/fmicb.2020.01017

Figure Lengend Snippet: Upregulated efflux pumps and differentially regulated transcription factors in the Δ dnm1 mgm1 tetOn mutant. Fifty-two ABC transporters, 211 MFS transporters and 312 transcription factors were identified in the reference genome of A. fumigatus . The identified genes were subsequently compared to the list of genes upregulated ( n = 612) or downregulated ( n = 628) in the Δ dnm1 mgm1 tetOn mutant. The number of upregulated ABC transporters (A) , upregulated MFS transporters (B) , and up- or downregulated transcription factors (C) are indicated in the intersections of the Venn diagrams. (D,E) Volcano plots of the expressed ABC transporters (D) and transcription factors (E) . The log 2 fold changes of the individual genes (x-axis) were plotted against the corresponding adjusted p -values (-log 10 ; y-axis). Differentially expressed genes ( p = 0.007) are indicated in orange. Genes that were further analyzed by constructing conditional mutants thereof were labeled accordingly.

Article Snippet: For identification of the ABC and MFS transporters as well as the transcription factors which are up- and downregulated in Δ dnm1 mgm1 tetOn the differentially expressed genes were compared with the genes coding the indicated proteins using the free software Knime .

Techniques: Mutagenesis, Labeling

Journal: Frontiers in Microbiology

Article Title: Differentially Regulated Transcription Factors and ABC Transporters in a Mitochondrial Dynamics Mutant Can Alter Azole Susceptibility of Aspergillus fumigatus

doi: 10.3389/fmicb.2020.01017

Figure Lengend Snippet: Deletion of genes encoding individual ABC transporters ( A ; abc1 , mdr1 ( abc2 ), abc3 and abc4 ) or transcription factors ( B ; mdu2, mdu3, mdu11, mdd3, gliZ ( mdd4 ) and mdd5 ) in the Δ dnm1 mgm1 tetOn mutant does not affect azole susceptibility. In a series of 10-fold dilutions derived from a starting suspension of 5 × 10 7 conidia ml – 1 of the indicated strains, aliquots of 3 μl were spotted onto Sabouraud (SAB) agar plates. When indicated (+ 0.41 μg ml – 1 Vori), medium was supplemented with voriconazole. Plates without voriconazole were incubated for 24 h at 37°C and plates containing voriconazole were incubated for 36 h at 37°C.

Article Snippet: For identification of the ABC and MFS transporters as well as the transcription factors which are up- and downregulated in Δ dnm1 mgm1 tetOn the differentially expressed genes were compared with the genes coding the indicated proteins using the free software Knime .

Techniques: Mutagenesis, Derivative Assay, Suspension, Incubation

Journal: Frontiers in Microbiology

Article Title: Differentially Regulated Transcription Factors and ABC Transporters in a Mitochondrial Dynamics Mutant Can Alter Azole Susceptibility of Aspergillus fumigatus

doi: 10.3389/fmicb.2020.01017

Figure Lengend Snippet: Possible mechanisms that mediate the increased azole tolerance of mitochondrial fission/fusion mutants. Mitochondrial dysfunction triggers altered expression of multiple azole tolerance-affecting genes (black arrows). Among the upregulated genes is the lanosterol 14α-demethylase Cyp51A which is the target enzyme that is inhibited by azole antifungals. Increased Cyp51A expression is a well-characterized azole resistance mechanism. Besides this, multiple transcription factors are up- and downregulated in the Δ dnm1 mgm1 tetOn mutant. At least five of these transcription factors are able to increase and two are able to decrease the azole susceptibility of A. fumigatus . In addition, multiple ABC and MFS transporters are upregulated in the Δ dnm1 mgm1 tetOn mutant. At least four of these ABC transporters are able to increase the azole tolerance of the mold. Deletion of individual identified transcription factors and ABC transporters does not diminish the increased azole tolerance of the Δ dnm1 mgm1 tetOn mutant. This suggests that the identified factors are functionally redundant or that other mechanisms are responsible for the azole tolerance of the mitochondrial fission/fusion mutant.

Article Snippet: For identification of the ABC and MFS transporters as well as the transcription factors which are up- and downregulated in Δ dnm1 mgm1 tetOn the differentially expressed genes were compared with the genes coding the indicated proteins using the free software Knime .

Techniques: Expressing, Mutagenesis