Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity
Fungal Oligopeptide Transporters (Fot) Fot1, Fot2 and Fot3 have been found in <i>Saccharomyces cerevisiae</i> wine strains, but not in strains from other environments. In the <i>S. cerevisiae</i> wine strain EC1118, Fot1 and Fot2 are responsible for a broader range of oligope...
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| Format: | Article |
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MDPI AG
2021-11-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/7/11/963 |
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| author | Carmen Becerra-Rodríguez Géraldine Taghouti Perrine Portier Sylvie Dequin Margarida Casal Sandra Paiva Virginie Galeote |
| author_facet | Carmen Becerra-Rodríguez Géraldine Taghouti Perrine Portier Sylvie Dequin Margarida Casal Sandra Paiva Virginie Galeote |
| author_sort | Carmen Becerra-Rodríguez |
| collection | DOAJ |
| description | Fungal Oligopeptide Transporters (Fot) Fot1, Fot2 and Fot3 have been found in <i>Saccharomyces cerevisiae</i> wine strains, but not in strains from other environments. In the <i>S. cerevisiae</i> wine strain EC1118, Fot1 and Fot2 are responsible for a broader range of oligopeptide utilization in comparison with strains not containing any Fot. This leads to better fermentation efficiency and an increased production of desirable organoleptic compounds in wine. Despite the benefits associated with Fot activity in <i>S. cerevisiae</i> within the wine environment, little is known about this family of transporters in yeast. The presence of Fot1, Fot2 and Fot3 in <i>S. cerevisiae</i> wine strains is due to horizontal gene transfer from the yeast <i>Torulaspora microellipsoides</i>, which harbors Fot2Tm, FotX and FotY proteins. Sequence analyses revealed that Fot family members have a high sequence identity in these yeast species. In this work, we aimed to further characterize the different Fot family members in terms of subcellular localization, gene expression in enological fermentation and substrate specificity. Using CRISPR/Cas9, we constructed <i>S. cerevisiae</i> wine strains containing each different Fot as the sole oligopeptide transporter to analyze their oligopeptide preferences by phenotype microarrays. The results of oligopeptide consumption show that Fot counterparts have different di-/tripeptide specificities, suggesting that punctual sequence divergence between <i>FOT</i> genes can be crucial for substrate recognition, binding and transport activity. <i>FOT</i> gene expression levels in different <i>S. cerevisiae</i> wine strains during enological fermentation, together with predicted binding motifs for transcriptional regulators in nitrogen metabolism, indicate that these transporters may be under the control of the Nitrogen Catabolite Repression (NCR) system. Finally, we demonstrated that Fot1 is located in the yeast plasma membrane. This work contributes to a better understanding of this family of oligopeptide transporters, which have demonstrated a key role in the utilization of oligopeptides by <i>S. cerevisiae</i> in enological fermentation. |
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| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-10T05:22:23Z |
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| spelling | doaj.art-2337340232ff491695529fd16d5f0ec62023-11-22T23:56:19ZengMDPI AGJournal of Fungi2309-608X2021-11-0171196310.3390/jof7110963Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence IdentityCarmen Becerra-Rodríguez0Géraldine Taghouti1Perrine Portier2Sylvie Dequin3Margarida Casal4Sandra Paiva5Virginie Galeote6SPO, Univ. Montpellier, INRAE, Institut Agro, F-34060 Montpellier, FranceUniv. Angers, Institut Agro, INRAE, IRHS, SFR QUASAV, CIRM-CFBP, F-49000 Angers, FranceUniv. Angers, Institut Agro, INRAE, IRHS, SFR QUASAV, CIRM-CFBP, F-49000 Angers, FranceSPO, Univ. Montpellier, INRAE, Institut Agro, F-34060 Montpellier, FranceCentre of Environmental and Molecular Biology, Department of Biology, Campus of Gualtar, University of Minho, 4710-057 Braga, PortugalCentre of Environmental and Molecular Biology, Department of Biology, Campus of Gualtar, University of Minho, 4710-057 Braga, PortugalSPO, Univ. Montpellier, INRAE, Institut Agro, F-34060 Montpellier, FranceFungal Oligopeptide Transporters (Fot) Fot1, Fot2 and Fot3 have been found in <i>Saccharomyces cerevisiae</i> wine strains, but not in strains from other environments. In the <i>S. cerevisiae</i> wine strain EC1118, Fot1 and Fot2 are responsible for a broader range of oligopeptide utilization in comparison with strains not containing any Fot. This leads to better fermentation efficiency and an increased production of desirable organoleptic compounds in wine. Despite the benefits associated with Fot activity in <i>S. cerevisiae</i> within the wine environment, little is known about this family of transporters in yeast. The presence of Fot1, Fot2 and Fot3 in <i>S. cerevisiae</i> wine strains is due to horizontal gene transfer from the yeast <i>Torulaspora microellipsoides</i>, which harbors Fot2Tm, FotX and FotY proteins. Sequence analyses revealed that Fot family members have a high sequence identity in these yeast species. In this work, we aimed to further characterize the different Fot family members in terms of subcellular localization, gene expression in enological fermentation and substrate specificity. Using CRISPR/Cas9, we constructed <i>S. cerevisiae</i> wine strains containing each different Fot as the sole oligopeptide transporter to analyze their oligopeptide preferences by phenotype microarrays. The results of oligopeptide consumption show that Fot counterparts have different di-/tripeptide specificities, suggesting that punctual sequence divergence between <i>FOT</i> genes can be crucial for substrate recognition, binding and transport activity. <i>FOT</i> gene expression levels in different <i>S. cerevisiae</i> wine strains during enological fermentation, together with predicted binding motifs for transcriptional regulators in nitrogen metabolism, indicate that these transporters may be under the control of the Nitrogen Catabolite Repression (NCR) system. Finally, we demonstrated that Fot1 is located in the yeast plasma membrane. This work contributes to a better understanding of this family of oligopeptide transporters, which have demonstrated a key role in the utilization of oligopeptides by <i>S. cerevisiae</i> in enological fermentation.https://www.mdpi.com/2309-608X/7/11/963Fungal Oligopeptide Transporters (Fot)oligopeptide transport<i>Saccharomyces cerevisiae</i>phenotype microarraysGFP labeling |
| spellingShingle | Carmen Becerra-Rodríguez Géraldine Taghouti Perrine Portier Sylvie Dequin Margarida Casal Sandra Paiva Virginie Galeote Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity Journal of Fungi Fungal Oligopeptide Transporters (Fot) oligopeptide transport <i>Saccharomyces cerevisiae</i> phenotype microarrays GFP labeling |
| title | Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity |
| title_full | Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity |
| title_fullStr | Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity |
| title_full_unstemmed | Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity |
| title_short | Yeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity |
| title_sort | yeast plasma membrane fungal oligopeptide transporters display distinct substrate preferences despite their high sequence identity |
| topic | Fungal Oligopeptide Transporters (Fot) oligopeptide transport <i>Saccharomyces cerevisiae</i> phenotype microarrays GFP labeling |
| url | https://www.mdpi.com/2309-608X/7/11/963 |
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