title: A stalled retrotranslocation complex reveals physical linkage between substrate recognition and proteasomal degradation during ER-associated degradation.
abstract: During endoplasmic reticulum-associated degradation (ERAD), misfolded lumenal and membrane proteins in the ER are recognized by the transmembrane Hrd1 ubiquitin ligase complex and retrotranslocated to the cytosol for ubiquitination and degradation. Although substrates are believed to be delivered to the proteasome only after the ATPase Cdc48p/p97 acts, there is limited knowledge about how the Hrd1 complex coordinates with Cdc48p/p97 and the proteasome to orchestrate substrate recognition and degradation. Here we provide evidence that inactivation of Cdc48p/p97 stalls retrotranslocation and triggers formation of a complex that contains the 26S proteasome, Cdc48p/p97, ubiquitinated substrates, select components of the Hrd1 complex, and the lumenal recognition factor, Yos9p. We propose that the actions of Cdc48p/p97 and the proteasome are tightly coupled during ERAD. Our data also support a model in which the Hrd1 complex links substrate recognition and degradation on opposite sides of the ER membrane.
authors.affiliation: Division of Biological Sciences, Graduate School of Science, Nagoya University, Nagoya, Aichi 464-8602 Japan. z47875a@cc.nagoya-u.ac.jp
authors.forename: Kunio
authors.initials: K
authors.lastname: Nakatsukasa
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Brodsky
authors.forename: Takumi
authors.initials: T
authors.lastname: Kamura
mesh_terms: D000251:Adenosine Triphosphatases / Q000172:deficiency / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000172:deficiency / Q000235:genetics*; D003593:Cytoplasm / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000235:genetics* / Q000378:metabolism; D060746:Endoplasmic Reticulum-Associated Degradation; D015966:Gene Expression Regulation, Fungal; D007425:Intracellular Membranes / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D017510:Protein Folding; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000378:metabolism; D054875:Ubiquitination; D000074405:Valosin Containing Protein
chemical_list: D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; C495027:Yos9 protein, S cerevisiae; C104621:HRD1 protein, S cerevisiae; D044767:Ubiquitin-Protein Ligases; D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases; C000614177:CDC48 protein, S cerevisiae; D000074405:Valosin Containing Protein
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