title: The E3 ubiquitin ligase parkin is recruited to the 26 S proteasome via the proteasomal ubiquitin receptor Rpn13.
abstract: Mutations in the Park2 gene, encoding the RING-HECT hybrid E3 ubiquitin ligase parkin, are responsible for a common familial form of Parkinson disease. By mono- and polyubiquitinating target proteins, parkin regulates various cellular processes, including degradation of proteins within the 26 S proteasome, a large multimeric degradation machine. In our attempt to further elucidate the function of parkin, we have identified the proteasomal ubiquitin receptor Rpn13/ADRM1 as a parkin-interacting protein. We show that the N-terminal ubiquitin-like (Ubl) domain of parkin binds directly to the pleckstrin-like receptor for ubiquitin (Pru) domain within Rpn13. Using mutational analysis and NMR, we find that Pru binding involves the hydrophobic patch surrounding Ile-44 in the parkin Ubl, a region that is highly conserved between ubiquitin and Ubl domains. However, compared with ubiquitin, the parkin Ubl exhibits greater than 10-fold higher affinity for the Pru domain. Moreover, knockdown of Rpn13 in cells increases parkin levels and abrogates parkin recruitment to the 26 S proteasome, establishing Rpn13 as the major proteasomal receptor for parkin. In contrast, silencing Rpn13 did not impair parkin recruitment to mitochondria or parkin-mediated mitophagy upon carbonyl cyanide m-chlorophenyl hydrazone-induced mitochondrial depolarization. However, it did delay the clearance of mitochondrial proteins (TIM23, TIM44, and TOM20) and enhance parkin autoubiquitination. Taken together, these findings implicate Rpn13 in linking parkin to the 26 S proteasome and regulating the clearance of mitochondrial proteins during mitophagy.
authors: From the McGill Parkinson Program and Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec H3A 2B4, Canada.
authors: Miguel A
authors: MA
authors: Aguileta
authors: the School of Medicine, University of Split, 21000 Split, Croatia.
authors: Jelena
authors: J
authors: Korac
authors: From the McGill Parkinson Program and Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec H3A 2B4, Canada.
authors: Thomas M
authors: TM
authors: Durcan
authors: the Department of Biochemistry, McGill University, Montreal, Quebec H3G 0B1, Canada.
authors: Jean-François
authors: JF
authors: Trempe
authors: From the McGill Parkinson Program and Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec H3A 2B4, Canada.
authors: Michael
authors: M
authors: Haber
authors: the Department of Biochemistry, McGill University, Montreal, Quebec H3G 0B1, Canada.
authors: Kalle
authors: K
authors: Gehring
authors: the Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, and.
authors: Suzanne
authors: S
authors: Elsasser
authors: Goethe University Medical School, D-60590 Frankfurt am Main, Germany.
authors: Oliver
authors: O
authors: Waidmann
authors: From the McGill Parkinson Program and Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec H3A 2B4, Canada.
authors: Edward A
authors: EA
authors: Fon
authors: Goethe University Medical School, D-60590 Frankfurt am Main, Germany husnjak@biochem2.de.
authors: Koraljka
authors: K
authors: Husnjak
mesh_terms: D001483:Base Sequence; D057809:HEK293 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D009682:Magnetic Resonance Spectroscopy; D008562:Membrane Glycoproteins / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
chemical_list: C500781:ADRM1 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease
keywords: E3 Ubiquitin Ligase; Parkin; Parkinson Disease; Proteasome; Rpn13; Ubiquitin; Ubiquitylation (Ubiquitination)