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Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex
Targeted endocytosis of plasma membrane (PM) proteins allows cells to adjust their complement of membrane proteins to changing extracellular conditions. For a wide variety of PM proteins, initiation of endocytosis is triggered by ubiquitination. In yeast, arrestin-related trafficking adaptors (ARTs)...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5156545/ https://www.ncbi.nlm.nih.gov/pubmed/27798240 http://dx.doi.org/10.1091/mbc.E16-08-0570 |
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author | Guiney, Evan L. Klecker, Till Emr, Scott D. |
author_facet | Guiney, Evan L. Klecker, Till Emr, Scott D. |
author_sort | Guiney, Evan L. |
collection | PubMed |
description | Targeted endocytosis of plasma membrane (PM) proteins allows cells to adjust their complement of membrane proteins to changing extracellular conditions. For a wide variety of PM proteins, initiation of endocytosis is triggered by ubiquitination. In yeast, arrestin-related trafficking adaptors (ARTs) enable a single ubiquitin ligase, Rsp5, to specifically and selectively target a wide range of PM proteins for ubiquitination and endocytosis. However, the mechanisms that allow ARTs to specifically recognize their appropriate substrates are unknown. We present the molecular features in the methionine permease Mup1 that are required for Art1-Rsp5–mediated ubiquitination and endocytosis. A combination of genetics, fluorescence microscopy, and biochemistry reveals three critical features that comprise an ART sorting signal in the Mup1 N-terminal cytosolic tail: 1) an extended acidic patch, 2) in close proximity to the first Mup1 transmembrane domain, and 3) close to the ubiquitinated lysines. We show that a functionally similar ART sorting signal is also required for the endocytosis of a second Art1-dependent cargo, Can1, suggesting a common mechanism for recognition of Art1 substrates. We isolate two separate suppressor mutations in the Art1 C-terminal domain that allele-specifically restore endocytosis of two Mup1 acidic patch mutants, consistent with an interaction between the Art1 C-terminus and the Mup1 acidic patch. We propose that this interaction is required for recruitment of the Art1-Rsp5 ubiquitination complex. |
format | Online Article Text |
id | pubmed-5156545 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-51565452017-03-02 Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex Guiney, Evan L. Klecker, Till Emr, Scott D. Mol Biol Cell Articles Targeted endocytosis of plasma membrane (PM) proteins allows cells to adjust their complement of membrane proteins to changing extracellular conditions. For a wide variety of PM proteins, initiation of endocytosis is triggered by ubiquitination. In yeast, arrestin-related trafficking adaptors (ARTs) enable a single ubiquitin ligase, Rsp5, to specifically and selectively target a wide range of PM proteins for ubiquitination and endocytosis. However, the mechanisms that allow ARTs to specifically recognize their appropriate substrates are unknown. We present the molecular features in the methionine permease Mup1 that are required for Art1-Rsp5–mediated ubiquitination and endocytosis. A combination of genetics, fluorescence microscopy, and biochemistry reveals three critical features that comprise an ART sorting signal in the Mup1 N-terminal cytosolic tail: 1) an extended acidic patch, 2) in close proximity to the first Mup1 transmembrane domain, and 3) close to the ubiquitinated lysines. We show that a functionally similar ART sorting signal is also required for the endocytosis of a second Art1-dependent cargo, Can1, suggesting a common mechanism for recognition of Art1 substrates. We isolate two separate suppressor mutations in the Art1 C-terminal domain that allele-specifically restore endocytosis of two Mup1 acidic patch mutants, consistent with an interaction between the Art1 C-terminus and the Mup1 acidic patch. We propose that this interaction is required for recruitment of the Art1-Rsp5 ubiquitination complex. The American Society for Cell Biology 2016-12-15 /pmc/articles/PMC5156545/ /pubmed/27798240 http://dx.doi.org/10.1091/mbc.E16-08-0570 Text en © 2016 Guiney, Klecker, and Emr. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License(http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Guiney, Evan L. Klecker, Till Emr, Scott D. Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex |
title | Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex |
title_full | Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex |
title_fullStr | Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex |
title_full_unstemmed | Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex |
title_short | Identification of the endocytic sorting signal recognized by the Art1-Rsp5 ubiquitin ligase complex |
title_sort | identification of the endocytic sorting signal recognized by the art1-rsp5 ubiquitin ligase complex |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5156545/ https://www.ncbi.nlm.nih.gov/pubmed/27798240 http://dx.doi.org/10.1091/mbc.E16-08-0570 |
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