Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans

Evangelinos, Minoas; Anagnostopoulos, Gerasimos; Karvela-Kalogeraki, Iliana; Stathopoulou, Panagiota; Scazzocchio, Claudio; Diallinas, George

doi:10.1016/j.fgb.2015.05.007

Cited by 9 publications

(19 citation statements)

References 60 publications

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“…A. Northern blot analysis of nmeA steady state mRNA levels in strains carrying the transposon element AnMinos (Evangelinos et al ., ) . AnMinos yA ‐ stands for the original strain carrying the AnMinos integrated in the niaD genomic locus, whereas Fur‐1 and Fur‐2 correspond to isogenic strains with transposition events of AnMinos in the 5' upstream region of nmeA , as shown in Supporting Information Figure S1.…”

Section: Resultsmentioning

confidence: 97%

“…We have previously isolated three A. nidulans mutants (Fur‐1, −2 and −3) resistant to 5‐FU arising from three distinct transpositions of the An Minos transposable elements in the 5' upstream region of the functionally uncharacterized gene AN2746 (Evangelinos et al ., ; Supporting Information Fig. S1, upper panel).…”

Section: Resultsmentioning

confidence: 99%

“…Fur-1, which was significantly closer to the predicted translation start of AN2746 (-264 nt) conferred relatively the strongest resistance to the fluorinated nucleobase/nucleoside analogues (Fur-1 > Fur-2 or Fur-3). In silico sequence analysis of the AN2746 open reading frame (ORF) showed that it encodes for a putative MFS transporter that exhibits low sequence similarity with putative members of a monocarboxylate transporter family (MCTs) (Evangelinos et al, 2015).…”

Section: Resultsmentioning

confidence: 99%

“…Contrastingly, the gpdA p -nmeA strain showed a sensitive phenotype, similar to wild-type, on itraconazole, Hygromycin D, acetic acid, propionic acid, 3-bromopyruvate, amino acid toxic analogues (D-serine and p-fluorophenylalanine), spermidine, spermine, putrescine, hydrogen peroxide, methylene blue, ethanol A. Northern blot analysis of nmeA steady state mRNA levels in strains carrying the transposon element AnMinos (Evangelinos et al, 2015). AnMinos yAstands for the original strain carrying the AnMinos integrated in the niaD genomic locus, whereas Fur-1 and Fur-2 correspond to isogenic strains with transposition events of AnMinos in the 5' upstream region of nmeA, as shown in Supporting Information Figure S1.…”

Section: Nmea Is a Very Low-transcribed Gene Under Standard Laboratormentioning

confidence: 94%

“…While rational reverse genetic approaches for protein functional analysis concern the generation of knock-outs, transposon-based analysis is based on the generation of mutant phenotypes, which allows the identification of genes through their hyper-expression (hypermorphs) or down-regulation of expression (hypomorphs), rather than complete knock-out. Recently, we developed an efficient novel transposon mutagenesis approach, based on the Minos element of Drosophila hydeii (Pavlopoulos et al, 2007) that allowed us to obtain morphological or drugresistant mutants in the model filamentous fungus Aspergillus nidulans (Evangelinos et al, 2015). In our screens using AnMinos, we obtained evidence that a number of morphological mutants isolated were due to hyper-expression of genes located next to transposon insertions.…”

Section: Introductionmentioning

confidence: 99%

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NmeA, a novel efflux transporter specific for nucleobases and nucleosides, contributes to metal resistance in Aspergillus nidulans

Balaska

Myrianthopoulos

Tselika

et al. 2017

Molecular Microbiology

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Through Minos transposon mutagenesis we obtained A. nidulans mutants resistant to 5-fluorouracil due to insertions into the upstream region of the uncharacterized gene nmeA, encoding a Major Facilitator Superfamily (MFS) transporter. Minos transpositions increased nmeA transcription, which is otherwise extremely low under all conditions tested. To dissect the function of NmeA we used strains overexpressing or genetically lacking the nmeA gene. Strains overexpressing NmeA are resistant to toxic purine analogues, but also, to cadmium, zinc and borate, whereas an isogenic nmeAΔ null mutant exhibits increased sensitivity to these compounds. We provide direct evidence that nmeA overexpression leads to efflux of adenine, xanthine, uric acid and allantoin, the latter two being intermediate metabolites of purine catabolism that are toxic when accumulated cytoplasmically due to relevant genetic lesions. By using a functional GFP-tagged version we show that NmeA is a plasma membrane transporter. Homology modeling and docking approaches identified a single purine binding site and a tentative substrate translocation trajectory in NmeA. Orthologues of NmeA are present in all Aspergilli and other Eurotiomycetes, but are absent from other fungi or non-fungal organisms. NmeA is thus the founding member of a new class of transporters essential for fungal success under specific toxic conditions.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Nmea Is a Very Low-transcribed Gene Under Standard Laboratormentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

NmeA, a novel efflux transporter specific for nucleobases and nucleosides, contributes to metal resistance in Aspergillus nidulans

Balaska

Myrianthopoulos

Tselika

et al. 2017

Molecular Microbiology

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An inducible tool for random mutagenesis in Aspergillus niger based on the transposon Vader

Paun

Nitsche

Homan

et al. 2016

Appl Microbiol Biotechnol

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The ascomycete Aspergillus niger is widely used in the biotechnology, for instance in producing most of the world's citric acid. It is also known as a major food and feed contaminant. While generation of gene knockouts for functional genomics has become feasible in ku70 mutants, analyzing gene functions or metabolic pathways remains a laborious task. An unbiased transposon-based mutagenesis approach may aid this process of analyzing gene functions by providing mutant libraries in a short time. The Vader transposon is a non-autonomous DNA-transposon, which is activated by the homologous tan1-transposase. However, in the most commonly used lab strain of A. niger (N400 strain and derivatives), we found that the transposase, encoded by the tan1 gene, is mutated and inactive. To establish a Vader transposon-based mutagenesis system in the N400 background, we expressed the functional transposase of A. niger strain CBS 513.88 under the control of an inducible promoter based on the Tet-on system, which is activated in the presence of the antibiotic doxycycline (DOX). Increasing amounts of doxycycline lead to higher Vader excision frequencies, whereas little to none activity of Vader was observed without addition of doxycycline. Hence, this system appears to be suitable for producing stable mutants in the A. niger N400 background.

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The crustacean model Parhyale hawaiensis

Paris

Wolff

Patel

et al. 2022

Current Topics in Developmental Biology

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Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans

Cited by 9 publications

References 60 publications

NmeA, a novel efflux transporter specific for nucleobases and nucleosides, contributes to metal resistance in Aspergillus nidulans

NmeA, a novel efflux transporter specific for nucleobases and nucleosides, contributes to metal resistance in Aspergillus nidulans

An inducible tool for random mutagenesis in Aspergillus niger based on the transposon Vader

The crustacean model Parhyale hawaiensis

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