Doa10 is a membrane protein retrotranslocase in ER-associated protein degradation
In endoplasmic reticulum-associated protein degradation (ERAD), membrane proteins are ubiquitinated, extracted from the membrane, and degraded by the proteasome. The cytosolic ATPase Cdc48 drives extraction by pulling on polyubiquitinated substrates. How hydrophobic transmembrane (TM) segments are m...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2020-06-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/56945 |
| _version_ | 1811199598120665088 |
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| author | Claudia C Schmidt Vedran Vasic Alexander Stein |
| author_facet | Claudia C Schmidt Vedran Vasic Alexander Stein |
| author_sort | Claudia C Schmidt |
| collection | DOAJ |
| description | In endoplasmic reticulum-associated protein degradation (ERAD), membrane proteins are ubiquitinated, extracted from the membrane, and degraded by the proteasome. The cytosolic ATPase Cdc48 drives extraction by pulling on polyubiquitinated substrates. How hydrophobic transmembrane (TM) segments are moved from the phospholipid bilayer into cytosol, often together with hydrophilic and folded ER luminal protein parts, is not known. Using a reconstituted system with purified proteins from Saccharomyces cerevisiae, we show that the ubiquitin ligase Doa10 (Teb-4/MARCH6 in animals) is a retrotranslocase that facilitates membrane protein extraction. A substrate’s TM segment interacts with the membrane-embedded domain of Doa10 and then passively moves into the aqueous phase. Luminal substrate segments cross the membrane in an unfolded state. Their unfolding occurs on the luminal side of the membrane by cytoplasmic Cdc48 action. Our results reveal how a membrane-bound retrotranslocase cooperates with the Cdc48 ATPase in membrane protein extraction. |
| first_indexed | 2024-04-12T01:51:25Z |
| format | Article |
| id | doaj.art-3a2835be509545ed9c2af3084896f6d2 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T01:51:25Z |
| publishDate | 2020-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-3a2835be509545ed9c2af3084896f6d22022-12-22T03:52:56ZengeLife Sciences Publications LtdeLife2050-084X2020-06-01910.7554/eLife.56945Doa10 is a membrane protein retrotranslocase in ER-associated protein degradationClaudia C Schmidt0https://orcid.org/0000-0002-7642-7081Vedran Vasic1https://orcid.org/0000-0003-2575-1006Alexander Stein2https://orcid.org/0000-0002-2696-0611Research Group Membrane Protein Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, GermanyResearch Group Membrane Protein Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, GermanyResearch Group Membrane Protein Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, GermanyIn endoplasmic reticulum-associated protein degradation (ERAD), membrane proteins are ubiquitinated, extracted from the membrane, and degraded by the proteasome. The cytosolic ATPase Cdc48 drives extraction by pulling on polyubiquitinated substrates. How hydrophobic transmembrane (TM) segments are moved from the phospholipid bilayer into cytosol, often together with hydrophilic and folded ER luminal protein parts, is not known. Using a reconstituted system with purified proteins from Saccharomyces cerevisiae, we show that the ubiquitin ligase Doa10 (Teb-4/MARCH6 in animals) is a retrotranslocase that facilitates membrane protein extraction. A substrate’s TM segment interacts with the membrane-embedded domain of Doa10 and then passively moves into the aqueous phase. Luminal substrate segments cross the membrane in an unfolded state. Their unfolding occurs on the luminal side of the membrane by cytoplasmic Cdc48 action. Our results reveal how a membrane-bound retrotranslocase cooperates with the Cdc48 ATPase in membrane protein extraction.https://elifesciences.org/articles/56945ERADprotein quality controlprotein translocationubiquitin proteasome systemmembrane proteinsin vitro reconstitution |
| spellingShingle | Claudia C Schmidt Vedran Vasic Alexander Stein Doa10 is a membrane protein retrotranslocase in ER-associated protein degradation eLife ERAD protein quality control protein translocation ubiquitin proteasome system membrane proteins in vitro reconstitution |
| title | Doa10 is a membrane protein retrotranslocase in ER-associated protein degradation |
| title_full | Doa10 is a membrane protein retrotranslocase in ER-associated protein degradation |
| title_fullStr | Doa10 is a membrane protein retrotranslocase in ER-associated protein degradation |
| title_full_unstemmed | Doa10 is a membrane protein retrotranslocase in ER-associated protein degradation |
| title_short | Doa10 is a membrane protein retrotranslocase in ER-associated protein degradation |
| title_sort | doa10 is a membrane protein retrotranslocase in er associated protein degradation |
| topic | ERAD protein quality control protein translocation ubiquitin proteasome system membrane proteins in vitro reconstitution |
| url | https://elifesciences.org/articles/56945 |
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