title: Proteasome 19S RP binding to the Sec61 channel plays a key role in ERAD.
abstract: Import of secretory proteins into the Endoplasmic Reticulum (ER) is an established function of the Sec61 channel. The contribution of the Sec61 channel to export of misfolded proteins from the ER for degradation by proteasomes is still controversial, but the proteasome 19S regulatory particle (RP) is necessary and sufficient for extraction of specific misfolded proteins from the ER, and binds directly to the Sec61 channel. In this work we have identified an import-competent sec61 mutant, S353C, carrying a point mutation in ER-lumenal loop 7 which reduces affinity of the cytoplasmic face of the Sec61 channel for the 19S RP. This indicates that the interaction between the 19S RP and the Sec61 channel is dependent on conformational changes in Sec61p hinging on loop 7. The sec61-S353C mutant had no measurable ER import defects and did not cause ER stress in intact cells, but reduced ER-export of a 19S RP-dependent misfolded protein when proteasomes were limiting in a cell-free assay. Our data suggest that the interaction between the 19S RP and the Sec61 channel is essential for the export of specific substrates from the ER to the cytosol for proteasomal degradation.
authors: Department of Microbiology, Faculty of Natural Sciences and Technology VIII, Saarland University, 66123, Saarbrücken, Germany.
authors: Marie-Luise
authors: ML
authors: Kaiser
authors: Department of Microbiology, Faculty of Natural Sciences and Technology VIII, Saarland University, 66123, Saarbrücken, Germany.
authors: Karin
authors: K
authors: Römisch
mesh_terms: D060746:Endoplasmic Reticulum-Associated Degradation; D026901:Membrane Transport Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D017354:Point Mutation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D000069816:SEC Translocation Channels; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
chemical_list: D026901:Membrane Transport Proteins; D000069816:SEC Translocation Channels; C499383:SEC61 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D046988:Proteasome Endopeptidase Complex; C119011:26S proteasome non-ATPase regulatory subunit 13