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Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation
The methylotrophic yeast Komagataella phaffii (Pichia pastoris) is homothallic and has been reported to switch mating type by an ancient inversion mechanism. Two mating-type (MAT) loci include homologs of the MATa and MATα transcription factor genes, with the expression from one locus downregulated...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5748462/ https://www.ncbi.nlm.nih.gov/pubmed/29061733 http://dx.doi.org/10.1128/MCB.00398-17 |
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author | Heistinger, Lina Gasser, Brigitte Mattanovich, Diethard |
author_facet | Heistinger, Lina Gasser, Brigitte Mattanovich, Diethard |
author_sort | Heistinger, Lina |
collection | PubMed |
description | The methylotrophic yeast Komagataella phaffii (Pichia pastoris) is homothallic and has been reported to switch mating type by an ancient inversion mechanism. Two mating-type (MAT) loci include homologs of the MATa and MATα transcription factor genes, with the expression from one locus downregulated by telomere position effects. However, not much is known about mating gene regulation, since the mixture of mating types complicates detailed investigations. In this study, we developed K. phaffii strains with stable mating types by deletion of the inverted-repeat region required for mating-type switching. These heterothallic strains retain their ability to mate with cells of the opposite mating type and were used to further elucidate mating gene regulation. Functional analysis of MAT mutant strains revealed the essential role of MATa2 and MATα1 in diploid cell formation. Disruption of MATa1 or MATα2 did not affect mating; however, in diploid cells, both genes are required for sporulation and the repression of shmoo formation. The heterothallic strains generated in this study allowed the first detailed characterization of mating gene regulation in K. phaffii. They will be a valuable tool for further studies investigating cell-type-specific behavior and will enable in-depth genetic analyses and strain hybridization in this industrially relevant yeast species. |
format | Online Article Text |
id | pubmed-5748462 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-57484622018-01-03 Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation Heistinger, Lina Gasser, Brigitte Mattanovich, Diethard Mol Cell Biol Research Article The methylotrophic yeast Komagataella phaffii (Pichia pastoris) is homothallic and has been reported to switch mating type by an ancient inversion mechanism. Two mating-type (MAT) loci include homologs of the MATa and MATα transcription factor genes, with the expression from one locus downregulated by telomere position effects. However, not much is known about mating gene regulation, since the mixture of mating types complicates detailed investigations. In this study, we developed K. phaffii strains with stable mating types by deletion of the inverted-repeat region required for mating-type switching. These heterothallic strains retain their ability to mate with cells of the opposite mating type and were used to further elucidate mating gene regulation. Functional analysis of MAT mutant strains revealed the essential role of MATa2 and MATα1 in diploid cell formation. Disruption of MATa1 or MATα2 did not affect mating; however, in diploid cells, both genes are required for sporulation and the repression of shmoo formation. The heterothallic strains generated in this study allowed the first detailed characterization of mating gene regulation in K. phaffii. They will be a valuable tool for further studies investigating cell-type-specific behavior and will enable in-depth genetic analyses and strain hybridization in this industrially relevant yeast species. American Society for Microbiology 2017-12-29 /pmc/articles/PMC5748462/ /pubmed/29061733 http://dx.doi.org/10.1128/MCB.00398-17 Text en Copyright © 2017 Heistinger et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Article Heistinger, Lina Gasser, Brigitte Mattanovich, Diethard Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation |
title | Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation |
title_full | Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation |
title_fullStr | Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation |
title_full_unstemmed | Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation |
title_short | Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation |
title_sort | creation of stable heterothallic strains of komagataella phaffii enables dissection of mating gene regulation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5748462/ https://www.ncbi.nlm.nih.gov/pubmed/29061733 http://dx.doi.org/10.1128/MCB.00398-17 |
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