TomA, the target of MAT1-1 in Aspergillus fumigatus, is required for cleistothecia formation presumably in a mating-type-specific manner
Pazen T, Ramsak B, Kueck U, Krappmann S (2022)
Publication Type: Conference contribution
Publication year: 2022
Journal
Publisher: WILEY
City/Town: HOBOKEN
Pages Range: 28-28
Conference Proceedings Title: MYCOSES
DOI: 10.1093/g3journal/jkaa012
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.
Authors with CRIS profile
Tobias Pazen
Institute of Microbiology – Clinical Microbiology, Immunology and Hygiene
Sven Krappmann
Professur für Klinische Mikrobiologie und Immunologie
Involved external institutions
Germany (DE)How to cite
APA:
Pazen, T., Ramsak, B., Kueck, U., & Krappmann, S. (2022). TomA, the target of MAT1-1 in Aspergillus fumigatus, is required for cleistothecia formation presumably in a mating-type-specific manner. In MYCOSES (pp. 28-28). HOBOKEN: WILEY.
MLA:
Pazen, Tobias, et al. "TomA, the target of MAT1-1 in Aspergillus fumigatus, is required for cleistothecia formation presumably in a mating-type-specific manner." Proceedings of the MYCOSES HOBOKEN: WILEY, 2022. 28-28.
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