The \(\textit {Penicillium chrysogenum tom1}\) gene a major target of transcription factor MAT1-1-1 encodes a nuclear protein involved in sporulation
- Fungal mating-type loci (\(\it MAT\)) encode transcription factors (TFs) MAT1-1-1 and MAT1-2-1, which govern sexual reproduction as well as other developmental processes. In \(\textit {Penicillium chrysogenum}\), the major producer of the beta-lactam antibiotic penicillin, a recent chromatin immunoprecipitation followed by sequencing (ChIP-seq) analysis identified 254 genes as direct targets of MAT1-1-1, many of which encode thus far uncharacterized proteins. Here, we characterized one of the major targets of MAT1-1-1, the \(\it tom1\) gene, which encodes a protein highly conserved within the group of Eurotiomycetes fungi. Using fluorescence microscopy, we demonstrated binding of MAT1-1-1 to the \(\it tom1\) promoter by reporter gene analysis. Extensive electrophoretic mobility shift assays (EMSAs) further showed that the promoter sequence of \(\it tom1\) is bound \(\textit {in vitro}\) by both MAT1-1-1 and MAT1-2-1. This indicated an interaction between the two TFs, which was verified by yeast two-hybrid analysis. The sequence of \(\it tom1\) carries a nuclear localization sequence, and indeed its nuclear localization was verified by fluorescence microscopy. The \(\textit {in vivo}\) function of \(\it tom1\) was investigated using \(\it tom1\) deletion strains, as well as a complementing strain where the wild-type \(\it tom1\) gene was reintroduced. We found a clear sporulation defect in the deletion strain, which became more evident when the fungi were grown at an elevated temperature of 31°C.
Author: | Barbara RamšakORCiDGND, Ulrich KückORCiDGND |
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URN: | urn:nbn:de:hbz:294-102270 |
DOI: | https://doi.org/10.3389/ffunb.2022.937023 |
Parent Title (English): | Frontiers in fungal biology |
Publisher: | Frontiers |
Place of publication: | Lausanne, Schweiz |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2023/10/17 |
Date of first Publication: | 2022/07/14 |
Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
Tag: | Open Access Fonds Mating-type transcription factor; Penicillium chrysogenum; electrophoretic mobility shift assay; fluorescence microscopy; sporulation; yeast two-hybrid analysis |
Volume: | 3 |
Issue: | Artikel 937023 |
First Page: | 937023-1 |
Last Page: | 937023-11 |
Note: | Article Processing Charge funded by the Deutsche Forschungsgemeinschaft (DFG) and the Open Access Publication Fund of Ruhr-Universität Bochum. |
Institutes/Facilities: | Fakultät für Biologie und Biotechnologie, Arbeitskreis Allgemeine und Molekulare Botanik |
Dewey Decimal Classification: | Naturwissenschaften und Mathematik / Biowissenschaften, Biologie, Biochemie |
open_access (DINI-Set): | open_access |
faculties: | Fakultät für Biologie und Biotechnologie |
Licence (English): | Creative Commons - CC BY 4.0 - Attribution 4.0 International |