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

Autor: Panagiota Stathopoulou, George Diallinas, Minoas Evangelinos, Gerasimos Anagnostopoulos, Iliana Karvela-Kalogeraki, Claudio Scazzocchio
Přispěvatelé: Faculty of Biology, University of Athens, Panepistimioupolis, University of Athens, Panepistimioupolis, Imperial College London, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Department of Microbiology, Imperial College, London, United Kingdom., Imperial College, London, United Kingdom, Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 )
Jazyk: angličtina
Rok vydání: 2015
Předmět:
Zdroj: Fungal Genetics and Biology
Fungal Genetics and Biology, 2015, 81, pp.1-11. ⟨10.1016/j.fgb.2015.05.007⟩
Fungal Genetics and Biology, Elsevier, 2015, 81, pp.1-11. 〈10.1016/j.fgb.2015.05.007〉
Fungal Genetics and Biology, Elsevier, 2015, 81, pp.1-11. ⟨10.1016/j.fgb.2015.05.007⟩
ISSN: 1087-1845
1096-0937
DOI: 10.1016/j.fgb.2015.05.007⟩
Popis: International audience; Transposons constitute powerful genetic tools for gene inactivation, exon or promoter trapping and genome analyses. The Minos element from Drosophila hydei, a Tc1/mariner-like transposon, has proved as a very efficient tool for heterologous transposition in several metazoa. In filamentous fungi, only a handful of fungal-specific transposable elements have been exploited as genetic tools, with the impala Tc1/mariner element from Fusarium oxysporum being the most successful. Here, we developed a two-component transposition system to manipulate Minos transposition in Aspergillus nidulans (AnMinos). Our system allows direct selection of transposition events based on re-activation of niaD, a gene necessary for growth on nitrate as a nitrogen source. On average, among 10(8) conidiospores, we obtain up to ∼0.8×10(2) transposition events leading to the expected revertant phenotype (niaD(+)), while ∼16% of excision events lead to AnMinos loss. Characterized excision footprints consisted of the four terminal bases of the transposon flanked by the TA target duplication and led to no major DNA rearrangements. AnMinos transposition depends on the presence of its homologous transposase. Its frequency was not significantly affected by temperature, UV irradiation or the transcription status of the original integration locus (niaD). Importantly, transposition is dependent on nkuA, encoding an enzyme essential for non-homologous end joining of DNA in double-strand break repair. AnMinos proved to be an efficient tool for functional analysis as it seems to transpose in different genomic loci positions in all chromosomes, including a high proportion of integration events within or close to genes. We have used Minos to obtain morphological and toxic analogue resistant mutants. Interestingly, among morphological mutants some seem to be due to Minos-elicited over-expression of specific genes, rather than gene inactivation.
Databáze: OpenAIRE
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