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Title: Direct visualization of degradation microcompartments at the ER membrane

Abstract

To promote the biochemical reactions of life, cells can compartmentalize molecular interaction partners together within separated non–membrane-bound regions. It is unknown whether this strategy is used to facilitate protein degradation at specific locations within the cell. Leveraging in situ cryo-electron tomography to image the native molecular landscape of the unicellular alga Chlamydomonas reinhardtii , we discovered that the cytosolic protein degradation machinery is concentrated within ∼200-nm foci that contact specialized patches of endoplasmic reticulum (ER) membrane away from the ER–Golgi interface. These non–membrane-bound microcompartments exclude ribosomes and consist of a core of densely clustered 26S proteasomes surrounded by a loose cloud of Cdc48. Active proteasomes in the microcompartments directly engage with putative substrate at the ER membrane, a function canonically assigned to Cdc48. Live-cell fluorescence microscopy revealed that the proteasome clusters are dynamic, with frequent assembly and fusion events. We propose that the microcompartments perform ER-associated degradation, colocalizing the degradation machinery at specific ER hot spots to enable efficient protein quality control.

Authors:
 [1];  [1];  [1];  [1];  [1];  [2]; ORCiD logo [3]; ORCiD logo [1];  [1]; ORCiD logo [1]
  1. Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany,
  2. Department of Structural Cell Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany,
  3. Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095
Publication Date:
Research Org.:
Univ. of California, Los Angeles, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1580556
Alternate Identifier(s):
OSTI ID: 1625041
Grant/Contract Number:  
FC02-02ER63421
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 117 Journal Issue: 2; Journal ID: ISSN 0027-8424
Publisher:
Proceedings of the National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
Science & Technology - Other Topics

Citation Formats

Albert, Sahradha, Wietrzynski, Wojciech, Lee, Chia-Wei, Schaffer, Miroslava, Beck, Florian, Schuller, Jan M., Salomé, Patrice A., Plitzko, Jürgen M., Baumeister, Wolfgang, and Engel, Benjamin D. Direct visualization of degradation microcompartments at the ER membrane. United States: N. p., 2019. Web. doi:10.1073/pnas.1905641117.
Albert, Sahradha, Wietrzynski, Wojciech, Lee, Chia-Wei, Schaffer, Miroslava, Beck, Florian, Schuller, Jan M., Salomé, Patrice A., Plitzko, Jürgen M., Baumeister, Wolfgang, & Engel, Benjamin D. Direct visualization of degradation microcompartments at the ER membrane. United States. https://doi.org/10.1073/pnas.1905641117
Albert, Sahradha, Wietrzynski, Wojciech, Lee, Chia-Wei, Schaffer, Miroslava, Beck, Florian, Schuller, Jan M., Salomé, Patrice A., Plitzko, Jürgen M., Baumeister, Wolfgang, and Engel, Benjamin D. Fri . "Direct visualization of degradation microcompartments at the ER membrane". United States. https://doi.org/10.1073/pnas.1905641117.
@article{osti_1580556,
title = {Direct visualization of degradation microcompartments at the ER membrane},
author = {Albert, Sahradha and Wietrzynski, Wojciech and Lee, Chia-Wei and Schaffer, Miroslava and Beck, Florian and Schuller, Jan M. and Salomé, Patrice A. and Plitzko, Jürgen M. and Baumeister, Wolfgang and Engel, Benjamin D.},
abstractNote = {To promote the biochemical reactions of life, cells can compartmentalize molecular interaction partners together within separated non–membrane-bound regions. It is unknown whether this strategy is used to facilitate protein degradation at specific locations within the cell. Leveraging in situ cryo-electron tomography to image the native molecular landscape of the unicellular alga Chlamydomonas reinhardtii , we discovered that the cytosolic protein degradation machinery is concentrated within ∼200-nm foci that contact specialized patches of endoplasmic reticulum (ER) membrane away from the ER–Golgi interface. These non–membrane-bound microcompartments exclude ribosomes and consist of a core of densely clustered 26S proteasomes surrounded by a loose cloud of Cdc48. Active proteasomes in the microcompartments directly engage with putative substrate at the ER membrane, a function canonically assigned to Cdc48. Live-cell fluorescence microscopy revealed that the proteasome clusters are dynamic, with frequent assembly and fusion events. We propose that the microcompartments perform ER-associated degradation, colocalizing the degradation machinery at specific ER hot spots to enable efficient protein quality control.},
doi = {10.1073/pnas.1905641117},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 2,
volume = 117,
place = {United States},
year = {Fri Dec 27 00:00:00 EST 2019},
month = {Fri Dec 27 00:00:00 EST 2019}
}

Journal Article:
Free Publicly Available Full Text
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https://doi.org/10.1073/pnas.1905641117

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