Characterization of the proteasome interaction with the Sec61 channel in the endoplasmic reticulum

• Published in: Journal of cell science Vol. 120; no. 4; pp. 682 - 691
• Main Authors: Ng, Waiyan, Sergeyenko, Tatiana, Zeng, Naiyan, Brown, Jeremy D, Römisch, Karin
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists Limited 15.02.2007
• Subjects: Adenosine Triphosphate - metabolism; Binding Sites; Endoplasmic Reticulum - metabolism; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - metabolism; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Membrane Transport Proteins; Models, Biological; Mutation; Proteasome Endopeptidase Complex - isolation & purification; Proteasome Endopeptidase Complex - metabolism; Protein Binding; Protein Structure, Tertiary; Ribosomes - metabolism; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; SEC Translocation Channels
• ISSN: 0021-9533; 1477-9137
• DOI: 10.1242/jcs.03351

Biogenesis of secretory proteins requires their translocation into the endoplasmic reticulum (ER) through the Sec61 channel. Proteins that fail to fold are transported back into the cytosol and are degraded by proteasomes. For many substrates this retrograde transport is affected by mutations in the Sec61 channel, and can be promoted by ATP and the 19S regulatory particle of the proteasome, which binds directly to the Sec61 channel via its base. Here, we identify mutations in SEC61 which reduce proteasome binding to the channel, and demonstrate that proteasomes and ribosomes bind differently to cytosolic domains of the channel. We found that Sec63p and BiP coprecipitate with ER-associated proteasomes, but Sec63p does not contribute to proteasome binding to the ER. The 19S base contains six AAA-ATPase subunits (Rpt proteins) that have non-equivalent functions in proteasome-mediated protein turnover and form a hetero-hexamer. Mutations in the ATP-binding sites of individual Rpt proteins all reduced the affinity of 19S complexes for the ER, suggesting that the 19S base in the ATP-bound conformation docks at the Sec61 channel.