Methanol and acriflavine resistance in Dictyostelium are caused by loss of catalase The GenBank accession number for the sequence reported in this paper is AF090443.

Garcia, Ma. Xenia U.; Roberts, Catherine; Alexander, Hannah; Stewart, Allison; Harwood, Adrian J.; Alexander, Stephen; Insall, Robert H.

doi:10.1099/00221287-148-1-333

Cited by 14 publications

(8 citation statements)

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“…Mutations which increase the tolerance to methanol in other organisms are so far reported to be mostly related to enzymes of methanol pathways and thereby reducing the accumulation of toxic downstream metabolites. For example the methanol tolerance of Dictyostelium increased significantly by loss of a catalase, which is supposed to be the main enzyme in this organism oxidizing methanol to formaldehyde [ 42 ]. Also methanol tolerance of the methylotroph bacterium Bacillus methanolicus is dependent on the activities of the methanol dehydrogenase Mdh and the enzymes 3-hexulose 6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), responsible for utilization of formaldehyde [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

Identification of two mutations increasing the methanol tolerance of Corynebacterium glutamicum

Leßmeier

Wendisch

2015

BMC Microbiol

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BackgroundMethanol is present in most ecosystems and may also occur in industrial applications, e.g. as an impurity of carbon sources such as technical glycerol. Methanol often inhibits growth of bacteria, thus, methanol tolerance may limit fermentative production processes.ResultsThe methanol tolerance of the amino acid producing soil bacterium Corynebacterium glutamicum was improved by experimental evolution in the presence of methanol. The resulting strain Tol1 exhibited significantly increased growth rates in the presence of up to 1 M methanol. However, neither transcriptional changes nor increased enzyme activities of the linear methanol oxidation pathway were observed, which was in accordance with the finding that tolerance to the downstream metabolites formaldehyde and formate was not improved. Genome sequence analysis of strain Tol1 revealed two point mutations potentially relevant to enhanced methanol tolerance: one leading to the amino acid exchange A165T of O-acetylhomoserine sulfhydrolase MetY and the other leading to shortened CoA transferase Cat (Q342*). Introduction of either mutation into the genome of C. glutamicum wild type increased methanol tolerance and introduction of both mutations into C. glutamicum was sufficient to achieve methanol tolerance almost indistinguishable from that of strain Tol1.ConclusionThe methanol tolerance of C. glutamicum can be increased by two point mutations leading to amino acid exchange of O-acetylhomoserine sulfhydrolase MetY and shortened CoA transferase Cat. Introduction of these mutations into producer strains may be helpful when using carbon sources containing methanol as component or impurity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0558-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Identification of two mutations increasing the methanol tolerance of Corynebacterium glutamicum

Leßmeier

Wendisch

2015

BMC Microbiol

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“…Acriflavine has been used as a topical antiseptic due to its antibiotic activity against fungi, bacteria, virus, and parasites [ 2 , 3 ]. It is also noteworthy that acriflavine is apparently oxidized in vivo to generate its active form [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

RNA-sequencing analysis of Trichophyton rubrumtranscriptome in response to sublethal doses of acriflavine

et al. 2014

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Background The dermatophyte Trichophyton rubrum is an anthropophilic filamentous fungus that infects keratinized tissues and is the most common etiologic agent isolated in human dermatophytoses. The clinical treatment of these infections is challenging because only few antifungal drugs are commercially available. To understand the mode of action of cytotoxic drugs against fungi, we evaluated the time-dependent effects of acriflavine on T. rubrum transcriptome using high-throughput RNA-sequencing (RNA-seq) technology. Results RNA-seq analysis generated approximately 200 million short reads that were mapped to the Broad Institute's Dermatophyte Comparative Database before differential gene expression analysis was performed. By employing a stringent cut-off threshold of −1.5 and 1.5 log 2 -fold changes in gene expression, a subset of 490 unique genes were found to be modulated in T. rubrum in response to acriflavine exposure. Among the selected genes, 69 genes were modulated at all exposure time points. Functional categorization indicated the putative involvement of these genes in various cellular processes such as oxidation-reduction reaction, transmembrane transport, and metal ion binding. Interestingly, genes putatively involved in the pathogenicity of dermatophytoses were down-regulated suggesting that this drug interferes with the virulence of T. rubrum . Moreover, we identified 159 novel putative transcripts in intergenic regions and two transcripts in intron regions of T. rubrum genome. Conclusion The results provide insights into the molecular events underlying the stress responses of T. rubrum to acriflavine, revealing that this drug interfered with important molecular events involved in the establishment and maintenance of fungal infection in the host. In addition, the identification of novel transcripts will further enable the improvement of gene annotation and open reading frame prediction of T. rubrum and other dermatophyte genomes.

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“…However, we measured mutagenesis frequency in Dictyostelium by assessing their resistance to methanol (Garcia et al, 2002; Podgorski and Deering, 1980). This mutagenic reporter system relies on the fact that methanol is toxic to Dictyostelium because of its conversion by the enzyme Catalase A (CatA) into toxic formic acid (Garcia et al, 2002). Inactivation of the catA gene leads to a failure to convert methanol into formic acid and hence confers resistance to this alcohol (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Then, the clonal population was lifted, and up to 5×10 6 viable amoebae were plated onto 3% methanol-containing SM-agar plates. After 6 days, the number of methanol-resistant colonies per plate was scored and plotted relative to the colony-forming efficiency on K. aerogenes plates (Garcia et al, 2002; Podgorski and Deering, 1980). Finally, methanol-resistant clones were grown in HL5 and the catA gene was cloned into pTOPO for sequencing.…”

Section: Methodsmentioning

confidence: 99%

Xpf suppresses the mutagenic consequences of phagocytosis in Dictyostelium

Pontel

Langenick

Rosado

et al. 2016

Journal of Cell Science

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As time passes, mutations accumulate in the genomes of all living organisms. These changes promote genetic diversity, but also precipitate ageing and the initiation of cancer. Food is a common source of mutagens, but little is known about how nutritional factors cause lasting genetic changes in the consuming organism. Here, we describe an unusual genetic interaction between DNA repair in the unicellular amoeba Dictyostelium discoideum and its natural bacterial food source. We found that Dictyostelium deficient in the DNA repair nuclease Xpf (xpf−) display a severe and specific growth defect when feeding on bacteria. Despite being proficient in the phagocytosis and digestion of bacteria, over time, xpf− Dictyostelium feeding on bacteria cease to grow and in many instances die. The Xpf nuclease activity is required for sustained growth using a bacterial food source. Furthermore, the ingestion of this food source leads to a striking accumulation of mutations in the genome of xpf− Dictyostelium. This work therefore establishes Dictyostelium as a model genetic system to dissect nutritional genotoxicity, providing insight into how phagocytosis can induce mutagenesis and compromise survival fitness.

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Methanol and acriflavine resistance in Dictyostelium are caused by loss of catalase The GenBank accession number for the sequence reported in this paper is AF090443.

Cited by 14 publications

References 37 publications

Identification of two mutations increasing the methanol tolerance of Corynebacterium glutamicum

Identification of two mutations increasing the methanol tolerance of Corynebacterium glutamicum

RNA-sequencing analysis of Trichophyton rubrumtranscriptome in response to sublethal doses of acriflavine

Xpf suppresses the mutagenic consequences of phagocytosis in Dictyostelium

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