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• W3132985874 abstract "Mating-type transcription factors are master regulators of sexually related signal transduction pathways in fungi; however, their recognition of specific DNA sequences from target genes is widely undetermined. Here, we identified and characterized the DNA-binding sequence of the MAT1-1-1 alpha-box domain transcription factor from the human pathogen Aspergillus fumigatus. In order to explore MAT1-1-1 DNA-binding targets, we used the previously reported MAT1-1-1 binding motif from Penicillium chrysogenum, in a bioinformatics approach. We identified 18 A. fumigatus genes carrying the MAT1.1 sequence in their upstream region, among them genes for the α-pheromone precursor (PpgA), G-protein-coupled pheromone receptor (PreA), and for TomA, an unidentified protein. To validate our prediction further, quantification of transcript levels showed a decrease in expression of ppgA, tomA, and others in a MAT1-1 deletion strain. For a functional analysis of the binding sites, truncated variants of the A. fumigatus MAT1-1-1 gene were introduced into Escherichia coli for heterologous expression. The yield of recombinant protein was further optimized for the AfMAT1-1-178-235 variant that harbors an extended alpha-box domain. AfMAT1-1-178-235 bound to a subset of the most strongly upregulated genes: ppgA, preA, and tomA. The DNA-binding specificity was confirmed by testing mutated binding sequences, as well as performing competition experiments with specific and non-specific sequences. Finally, equilibrium dissociation constants of 1.83 ± 0.1 and 1.45 ± 0.26 µM were determined for AfMAT1-1-178-235 and fusion protein GST-AfMAT1-1-178-235. Collectively, these findings provide further insights into AfMAT1-1-1-mediated gene expression and imply that alpha-box domain regulators from other members of Eurotiales control fungal development in a conserved manner." @default.
• W3132985874 created "2021-03-01" @default.
• W3132985874 creator A5021457266 @default.
• W3132985874 creator A5022044880 @default.
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• W3132985874 creator A5057941883 @default.
• W3132985874 creator A5079723478 @default.
• W3132985874 creator A5084307476 @default.
• W3132985874 date "2020-12-08" @default.
• W3132985874 modified "2023-10-16" @default.
• W3132985874 title "The master regulator MAT1-1-1 of fungal mating binds to its targets via a conserved motif in the human pathogen<i>Aspergillus fumigatus</i>" @default.
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• W3132985874 doi "https://doi.org/10.1093/g3journal/jkaa012" @default.
• W3132985874 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8022922" @default.
• W3132985874 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/33598704" @default.
• W3132985874 hasPublicationYear "2020" @default.
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• W3132985874 hasAuthorship W3132985874A5079723478 @default.

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