Plasma membrane (PM) transporters of the major facilitator superfamily (MFS) are essential for cell metabolism, growth and response to stress or drugs. In Saccharomyces cerevisiae, Jen1 is a monocarboxylate/H+ symporter that provides a model to dissect the molecular details underlying cellular expression, transport mechanism and turnover of MFS transporters. Here, we present evidence revealing novel roles of the cytosolic N- and C- termini of Jen1 in its biogenesis, PM stability and transport activity, using functional analyses of Jen1 truncations and chimeric constructs with UapA, an endocytosis-insensitive transporter of Aspergillus nidulans. Our results show that both N- and C-termini are critical for Jen1 trafficking to the PM, transport activity and endocytosis. Importantly, we provide evidence that Jen1 N- and C- termini undergo transport-dependent dynamic intra-molecular interactions, which affect the transport activity and turnover of Jen1. Our results support an emerging concept where the cytoplasmic termini of PM transporters control transporter cell-surface stability and function through flexible intra-molecular interactions with each other. These findings may be extended to other MFS members to understand conserved and evolving mechanisms underlying transporter structure-function relationships.
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RESEARCH ARTICLE|
19 April 2022
Interactions of cytosolic tails in the Jen1 carboxylate transporter are critical for trafficking and transport activity
Cláudia Barata-Antunes,
Cláudia Barata-Antunes
*
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
2
Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho
, Portugal
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Gabriel Talaia
,
Gabriel Talaia
*
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
3
Department of Cell Biology, Yale University School of Medicine, New Haven, CT
, USA
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George Broutzakis
,
George Broutzakis
4
Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis 15784, Athens
, Greece
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David Ribas,
David Ribas
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
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Pieter De Beule,
Pieter De Beule
5
International Iberian Nanotechnology Laboratory, Avenida Mestre José Veiga s/n, Braga
, Portugal
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Margarida Casal,
Margarida Casal
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
2
Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho
, Portugal
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Christopher J. Stefan
,
Christopher J. Stefan
6
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion
, Greece
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George Diallinas
,
4
Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis 15784, Athens
, Greece
6
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion
, Greece
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Sandra Paiva
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
2
Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho
, Portugal
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Cláudia Barata-Antunes
*
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
2
Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho
, Portugal
Gabriel Talaia
*
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
3
Department of Cell Biology, Yale University School of Medicine, New Haven, CT
, USA
George Broutzakis
4
Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis 15784, Athens
, Greece
David Ribas
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
Pieter De Beule
5
International Iberian Nanotechnology Laboratory, Avenida Mestre José Veiga s/n, Braga
, Portugal
Margarida Casal
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
2
Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho
, Portugal
Christopher J. Stefan
6
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion
, Greece
4
Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis 15784, Athens
, Greece
6
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion
, Greece
1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga
, Portugal
2
Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho
, Portugal
*
The authors contributed equally to the work
Received:
28 Mar 2022
Accepted:
05 Apr 2022
Online ISSN: 1477-9137
Print ISSN: 0021-9533
Funding Group:
- Award Group:
- Funder(s): Fundacao para a Cioncia e a Tecnologia (FCT) I.P.
- Award Id(s): UIDB/04050/2020
- Funder(s):
Funding Group:
- Award Group:
- Funder(s): European Regional Development Fund
- Award Id(s): PTDC/BIA-MIC/5246/2020
- Funder(s):
Funding Group:
- Award Group:
- Funder(s): Fondation Sant
- Award Id(s): KE17237
- Funder(s):
J Cell Sci jcs.260059.
Article history
Received:
28 Mar 2022
Accepted:
05 Apr 2022
Currently Viewing Accepted Manuscript - Newer Version Available
19 May 2022
Citation
Cláudia Barata-Antunes, Gabriel Talaia, George Broutzakis, David Ribas, Pieter De Beule, Margarida Casal, Christopher J. Stefan, George Diallinas, Sandra Paiva; Interactions of cytosolic tails in the Jen1 carboxylate transporter are critical for trafficking and transport activity. J Cell Sci 2022; jcs.260059. doi: https://doi.org/10.1242/jcs.260059
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