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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Autores principales: Carmen Becerra-Rodríguez, Géraldine Taghouti, Perrine Portier, Sylvie Dequin, Margarida Casal, Sandra Paiva, Virginie Galeote
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Publicado: MDPI AG 2021
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spelling oai:doaj.org-article:2337340232ff491695529fd16d5f0ec62021-11-25T18:06:13ZYeast Plasma Membrane Fungal Oligopeptide Transporters Display Distinct Substrate Preferences despite Their High Sequence Identity10.3390/jof71109632309-608Xhttps://doaj.org/article/2337340232ff491695529fd16d5f0ec62021-11-01T00:00:00Zhttps://www.mdpi.com/2309-608X/7/11/963https://doaj.org/toc/2309-608XFungal 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.Carmen Becerra-RodríguezGéraldine TaghoutiPerrine PortierSylvie DequinMargarida CasalSandra PaivaVirginie GaleoteMDPI AGarticleFungal Oligopeptide Transporters (Fot)oligopeptide transport<i>Saccharomyces cerevisiae</i>phenotype microarraysGFP labelingBiology (General)QH301-705.5ENJournal of Fungi, Vol 7, Iss 963, p 963 (2021)
institution DOAJ
collection DOAJ
language EN
topic Fungal Oligopeptide Transporters (Fot)
oligopeptide transport
<i>Saccharomyces cerevisiae</i>
phenotype microarrays
GFP labeling
Biology (General)
QH301-705.5
spellingShingle Fungal Oligopeptide Transporters (Fot)
oligopeptide transport
<i>Saccharomyces cerevisiae</i>
phenotype microarrays
GFP labeling
Biology (General)
QH301-705.5
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
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.
format article
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
title 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_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_sort yeast plasma membrane fungal oligopeptide transporters display distinct substrate preferences despite their high sequence identity
publisher MDPI AG
publishDate 2021
url https://doaj.org/article/2337340232ff491695529fd16d5f0ec6
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