ADD66, a gene involved in the endoplasmic reticulum-associated degradation of alpha-1-antitrypsin-Z in yeast, facilitates proteasome activity and assembly.
Antitrypsin deficiency is a primary cause of juvenile liver disease, and it arises from expression of the "Z" variant of the alpha-1 protease inhibitor (A1Pi). Whereas A1Pi is secreted from the liver, A1PiZ is retrotranslocated from the endoplasmic reticulum (ER) and degraded by the proteasome, an event that may offset liver damage. To better define the mechanism of A1PiZ degradation, a yeast expression system was developed previously, and a gene, ADD66, was identified that facilitates A1PiZ turnover. We report here that ADD66 encodes an approximately 30-kDa soluble, cytosolic protein and that the chymotrypsin-like activity of the proteasome is reduced in add66Delta mutants. This reduction in activity may arise from the accumulation of 20S proteasome assembly intermediates or from qualitative differences in assembled proteasomes. Add66p also seems to be a proteasome substrate. Consistent with its role in ER-associated degradation (ERAD), synthetic interactions are observed between the genes encoding Add66p and Ire1p, a transducer of the unfolded protein response, and yeast deleted for both ADD66 and/or IRE1 accumulate polyubiquitinated proteins. These data identify Add66p as a proteasome assembly chaperone (PAC), and they provide the first link between PAC activity and ERAD.
Scott
Craig M
CM
Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA.
Kruse
Kristina B
KB
Schmidt
Béla Z
BZ
Perlmutter
David H
DH
McCracken
Ardythe A
AA
Brodsky
Jeffrey L
JL
D002918:Chymotrypsin / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D017353:Gene Deletion; D005800:Genes, Fungal; D008562:Membrane Glycoproteins / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D017510:Protein Folding; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D012995:Solubility; D000515:alpha 1-Antitrypsin / Q000378:metabolism*
D008562:Membrane Glycoproteins; D011498:Protein Precursors; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D000515:alpha 1-Antitrypsin; C082863:IRE1 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases; D002918:Chymotrypsin; D046988:Proteasome Endopeptidase Complex