The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation
Selective endocytosis followed by degradation is a major mechanism for downregulating plasma membrane transporters in response to specific environmental cues. In <i>Saccharomyces cerevisiae,</i> this endocytosis is promoted by ubiquitylation catalyzed by the Rsp5 ubiquitin-ligase, target...
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| Format: | Article |
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MDPI AG
2021-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/19/10208 |
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| author | Amalia H. Megarioti Cecilia Primo George C. Kapetanakis Alexandros Athanasopoulos Vicky Sophianopoulou Bruno André Christos Gournas |
| author_facet | Amalia H. Megarioti Cecilia Primo George C. Kapetanakis Alexandros Athanasopoulos Vicky Sophianopoulou Bruno André Christos Gournas |
| author_sort | Amalia H. Megarioti |
| collection | DOAJ |
| description | Selective endocytosis followed by degradation is a major mechanism for downregulating plasma membrane transporters in response to specific environmental cues. In <i>Saccharomyces cerevisiae,</i> this endocytosis is promoted by ubiquitylation catalyzed by the Rsp5 ubiquitin-ligase, targeted to transporters via adaptors of the alpha-arrestin family. However, the molecular mechanisms of this targeting and their control according to conditions remain incompletely understood. In this work, we dissect the molecular mechanisms eliciting the endocytosis of Can1, the arginine permease, in response to cycloheximide-induced TORC1 hyperactivation. We show that cycloheximide promotes Rsp5-dependent Can1 ubiquitylation and endocytosis in a manner dependent on the Bul1/2 alpha-arrestins. Also crucial for this downregulation is a short acidic patch sequence in the N-terminus of Can1 likely acting as a binding site for Bul1/2. The previously reported inhibition by cycloheximide of transporter recycling, from the trans-Golgi network to the plasma membrane, seems to additionally contribute to efficient Can1 downregulation. Our results also indicate that, contrary to the previously described substrate-transport elicited Can1 endocytosis mediated by the Art1 alpha-arrestin, Bul1/2-mediated Can1 ubiquitylation occurs independently of the conformation of the transporter. This study provides further insights into how distinct alpha-arrestins control the ubiquitin-dependent downregulation of a specific amino acid transporter under different conditions. |
| first_indexed | 2024-03-10T07:00:44Z |
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| id | doaj.art-61f8ce47719b4b91acca4f1a55da1eeb |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T07:00:44Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-61f8ce47719b4b91acca4f1a55da1eeb2023-11-22T16:06:04ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-09-0122191020810.3390/ijms221910208The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-HyperactivationAmalia H. Megarioti0Cecilia Primo1George C. Kapetanakis2Alexandros Athanasopoulos3Vicky Sophianopoulou4Bruno André5Christos Gournas6Microbial Molecular Genetics Laboratory, Institute of Biosciences and Applications, National Centre for Scientific Research “Demokritos”, Patr. Grigoriou E & 27 Neapoleos St., 15341 Agia Paraskevi, GreeceMolecular Physiology of the Cell Laboratory, Université Libre de Bruxelles (ULB), IBMM, 6041 Gosselies, BelgiumMolecular Physiology of the Cell Laboratory, Université Libre de Bruxelles (ULB), IBMM, 6041 Gosselies, BelgiumMicrobial Molecular Genetics Laboratory, Institute of Biosciences and Applications, National Centre for Scientific Research “Demokritos”, Patr. Grigoriou E & 27 Neapoleos St., 15341 Agia Paraskevi, GreeceMicrobial Molecular Genetics Laboratory, Institute of Biosciences and Applications, National Centre for Scientific Research “Demokritos”, Patr. Grigoriou E & 27 Neapoleos St., 15341 Agia Paraskevi, GreeceMolecular Physiology of the Cell Laboratory, Université Libre de Bruxelles (ULB), IBMM, 6041 Gosselies, BelgiumMicrobial Molecular Genetics Laboratory, Institute of Biosciences and Applications, National Centre for Scientific Research “Demokritos”, Patr. Grigoriou E & 27 Neapoleos St., 15341 Agia Paraskevi, GreeceSelective endocytosis followed by degradation is a major mechanism for downregulating plasma membrane transporters in response to specific environmental cues. In <i>Saccharomyces cerevisiae,</i> this endocytosis is promoted by ubiquitylation catalyzed by the Rsp5 ubiquitin-ligase, targeted to transporters via adaptors of the alpha-arrestin family. However, the molecular mechanisms of this targeting and their control according to conditions remain incompletely understood. In this work, we dissect the molecular mechanisms eliciting the endocytosis of Can1, the arginine permease, in response to cycloheximide-induced TORC1 hyperactivation. We show that cycloheximide promotes Rsp5-dependent Can1 ubiquitylation and endocytosis in a manner dependent on the Bul1/2 alpha-arrestins. Also crucial for this downregulation is a short acidic patch sequence in the N-terminus of Can1 likely acting as a binding site for Bul1/2. The previously reported inhibition by cycloheximide of transporter recycling, from the trans-Golgi network to the plasma membrane, seems to additionally contribute to efficient Can1 downregulation. Our results also indicate that, contrary to the previously described substrate-transport elicited Can1 endocytosis mediated by the Art1 alpha-arrestin, Bul1/2-mediated Can1 ubiquitylation occurs independently of the conformation of the transporter. This study provides further insights into how distinct alpha-arrestins control the ubiquitin-dependent downregulation of a specific amino acid transporter under different conditions.https://www.mdpi.com/1422-0067/22/19/10208ubiquitinα-arrestinendocytosistransporterarginineTarget of Rapamycin Complex 1 |
| spellingShingle | Amalia H. Megarioti Cecilia Primo George C. Kapetanakis Alexandros Athanasopoulos Vicky Sophianopoulou Bruno André Christos Gournas The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation International Journal of Molecular Sciences ubiquitin α-arrestin endocytosis transporter arginine Target of Rapamycin Complex 1 |
| title | The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation |
| title_full | The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation |
| title_fullStr | The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation |
| title_full_unstemmed | The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation |
| title_short | The Bul1/2 Alpha-Arrestins Promote Ubiquitylation and Endocytosis of the Can1 Permease upon Cycloheximide-Induced TORC1-Hyperactivation |
| title_sort | bul1 2 alpha arrestins promote ubiquitylation and endocytosis of the can1 permease upon cycloheximide induced torc1 hyperactivation |
| topic | ubiquitin α-arrestin endocytosis transporter arginine Target of Rapamycin Complex 1 |
| url | https://www.mdpi.com/1422-0067/22/19/10208 |
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