beta-Subunit appendages promote 20S proteasome assembly by overcoming an Ump1-dependent checkpoint.
Proteasomes are responsible for most intracellular protein degradation in eukaryotes. The 20S proteasome comprises a dyad-symmetric stack of four heptameric rings made from 14 distinct subunits. How it assembles is not understood. Most subunits in the central pair of beta-subunit rings are synthesized in precursor form. Normally, the beta5 (Doa3) propeptide is essential for yeast proteasome biogenesis, but overproduction of beta7 (Pre4) bypasses this requirement. Bypass depends on a unique beta7 extension, which contacts the opposing beta ring. The resulting proteasomes appear normal but assemble inefficiently, facilitating identification of assembly intermediates. Assembly occurs stepwise into precursor dimers, and intermediates contain the Ump1 assembly factor and a novel complex, Pba1-Pba2. beta7 incorporation occurs late and is closely linked to the association of two half-proteasomes. We propose that dimerization is normally driven by the beta5 propeptide, an intramolecular chaperone, but beta7 addition overcomes an Ump1-dependent assembly checkpoint and stabilizes the precursor dimer.
Li
Xia
X
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8114, USA.
Kusmierczyk
Andrew R
AR
Wong
Peter
P
Emili
Andrew
A
Hochstrasser
Mark
M
D003546:Cysteine Endopeptidases / Q000096:biosynthesis; D019281:Dimerization; D006801:Humans; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D046988:Proteasome Endopeptidase Complex / Q000096:biosynthesis*; D011498:Protein Precursors / Q000096:biosynthesis*; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis*
D018832:Molecular Chaperones; D011498:Protein Precursors; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; C110856:proteasome maturation protein; D003546:Cysteine Endopeptidases; C080044:PRE2 protein, S cerevisiae; C478799:PRE4 protein, S cerevisiae; D046988:Proteasome Endopeptidase Complex