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Free keywords:
RETICULUM-ASSOCIATED DEGRADATION; UNFOLDED PROTEIN RESPONSE; CRYO-EM
STRUCTURE; ENDOPLASMIC-RETICULUM; PHASE-SEPARATION; CRYOELECTRON
TOMOGRAMS; LIQUID-LIKE; PROTEASOME; REVEALS; LOCALIZATIONproteasome; cdc48; ERAD; phase separation; cryo-electron tomography;
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 similar to 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 265 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.
