Structure of Rpn10 and its interactions with polyubiquitin chains and the proteasome subunit Rpn12.
Schizosaccharomyces pombe Rpn10 (SpRpn10) is a proteasomal ubiquitin (Ub) receptor located within the 19 S regulatory particle where it binds to subunits of both the base and lid subparticles. We have solved the structure of full-length SpRpn10 by determining the crystal structure of the von Willebrand factor type A domain and characterizing the full-length protein by NMR. We demonstrate that the single Ub-interacting motif (UIM) of SpRpn10 forms a 1:1 complex with Lys(48)-linked diUb, which it binds selectively over monoUb and Lys(63)-linked diUb. We further show that the SpRpn10 UIM binds to SpRpn12, a subunit of the lid subparticle, with an affinity comparable with Lys(48)-linked diUb. This is the first observation of a UIM binding other than a Ub fold and suggests that SpRpn12 could modulate the activity of SpRpn10 as a proteasomal Ub receptor.
Riedinger
Christiane
C
Laboratory of Molecular Biophysics, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom.
Boehringer
Jonas
J
Trempe
Jean-Francois
JF
Lowe
Edward D
ED
Brown
Nicholas R
NR
Gehring
Kalle
K
Noble
Martin E M
ME
Gordon
Colin
C
Endicott
Jane A
JA
D002151:Calorimetry / Q000379:methods; D002352:Carrier Proteins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D025821:Polyubiquitin / Q000737:chemistry*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011485:Protein Binding; D016601:RNA-Binding Proteins; D012568:Schizosaccharomyces / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D015398:Signal Transduction; D020349:Surface Plasmon Resonance; D025801:Ubiquitin / Q000737:chemistry; D014841:von Willebrand Factor / Q000737:chemistry
D002352:Carrier Proteins; D016601:RNA-Binding Proteins; C098743:RPN12 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D025801:Ubiquitin; C498507:pus1 protein, S pombe; D014841:von Willebrand Factor; D025821:Polyubiquitin; D046988:Proteasome Endopeptidase Complex; D008239:Lysine