title: Retrotranslocation of a misfolded luminal ER protein by the ubiquitin-ligase Hrd1p.
abstract: Misfolded, luminal endoplasmic reticulum (ER) proteins are retrotranslocated into the cytosol and degraded by the ubiquitin/proteasome system. This ERAD-L pathway requires a protein complex consisting of the ubiquitin ligase Hrd1p, which spans the ER membrane multiple times, and the membrane proteins Hrd3p, Usa1p, and Der1p. Here, we show that Hrd1p is the central membrane component in ERAD-L; its overexpression bypasses the need for the other components of the Hrd1p complex. Hrd1p function requires its oligomerization, which in wild-type cells is facilitated by Usa1p. Site-specific photocrosslinking indicates that, at early stages of retrotranslocation, Hrd1p interacts with a substrate segment close to the degradation signal. This interaction follows the delivery of substrate through other ERAD components, requires the presence of transmembrane segments of Hrd1p, and depends on both the ubiquitin ligase activity of Hrd1p and the function of the Cdc48p ATPase complex. Our results suggest a model for how Hrd1p promotes polypeptide movement through the ER membrane.
authors.affiliation: Howard Hughes Medical Institute and Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA. pedro.carvalho@crg.es
authors.forename: Pedro
authors.initials: P
authors.lastname: Carvalho
authors.forename: Ann Marie
authors.initials: AM
authors.lastname: Stanley
authors.forename: Tom A
authors.initials: TA
authors.lastname: Rapoport
mesh_terms: D004721:Endoplasmic Reticulum / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism; D017510:Protein Folding; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
chemical_list: D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins; C104621:HRD1 protein, S cerevisiae; D044767:Ubiquitin-Protein Ligases