Molecular architecture of the 26S proteasome holocomplex determined by an integrative approach.
The 26S proteasome is at the executive end of the ubiquitin-proteasome pathway for the controlled degradation of intracellular proteins. While the structure of its 20S core particle (CP) has been determined by X-ray crystallography, the structure of the 19S regulatory particle (RP), which recruits substrates, unfolds them, and translocates them to the CP for degradation, has remained elusive. Here, we describe the molecular architecture of the 26S holocomplex determined by an integrative approach based on data from cryoelectron microscopy, X-ray crystallography, residue-specific chemical cross-linking, and several proteomics techniques. The "lid" of the RP (consisting of Rpn3/5/6/7/8/9/11/12) is organized in a modular fashion. Rpn3/5/6/7/9/12 form a horseshoe-shaped heterohexamer, which connects to the CP and roofs the AAA-ATPase module, positioning the Rpn8/Rpn11 heterodimer close to its mouth. Rpn2 is rigid, supporting the lid, while Rpn1 is conformationally variable, positioned at the periphery of the ATPase ring. The ubiquitin receptors Rpn10 and Rpn13 are located in the distal part of the RP, indicating that they were recruited to the complex late in its evolution. The modular structure of the 26S proteasome provides insights into the sequence of events prior to the degradation of ubiquitylated substrates.
Department of Bioengineering and Therapeutic Sciences, California Institute of Quantitative Biosciences, 1700 4th Street, University of California, San Francisco, CA 94158, USA.
Keren
K
Lasker
Friedrich
F
Förster
Stefan
S
Bohn
Thomas
T
Walzthoeni
Elizabeth
E
Villa
Pia
P
Unverdorben
Florian
F
Beck
Ruedi
R
Aebersold
Andrej
A
Sali
Wolfgang
W
Baumeister
D020285:Cryoelectron Microscopy; D018360:Crystallography, X-Ray; D013058:Mass Spectrometry; D008958:Models, Molecular; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D040901:Proteomics; D012568:Schizosaccharomyces / Q000201:enzymology; D013379:Substrate Specificity
D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease