title: Structure of ubiquitylated-Rpn10 provides insight into its autoregulation mechanism.
abstract: Ubiquitin receptors decode ubiquitin signals into many cellular responses. Ubiquitin receptors also undergo coupled monoubiquitylation, and rapid deubiquitylation has hampered the characterization of the ubiquitylated state. Using bacteria that express a ubiquitylation apparatus, we purified and determined the crystal structure of the proteasomal ubiquitin-receptor Rpn10 in its ubiquitylated state. The structure shows a novel ubiquitin-binding patch that directs K84 ubiquitylation. Superimposition of ubiquitylated-Rpn10 onto electron-microscopy models of proteasomes indicates that the Rpn10-conjugated ubiquitin clashes with Rpn9, suggesting that ubiquitylation might be involved in releasing Rpn10 from the proteasome. Indeed, ubiquitylation on immobilized proteasomes dissociates the modified Rpn10 from the complex, while unmodified Rpn10 mainly remains associated. In vivo experiments indicate that contrary to wild type, Rpn10-K84R is stably associated with the proteasomal subunit Rpn9. Similarly Rpn10, but not ubiquitylated-Rpn10, binds Rpn9 in vitro. Thus we suggest that ubiquitylation functions to dissociate modified ubiquitin receptors from their targets, a function that promotes cyclic activity of ubiquitin receptors.
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Tal
authors.initials: T
authors.lastname: Keren-Kaplan
authors.affiliation: Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel.
authors.forename: Lee
authors.initials: L
authors.lastname: Zeev Peters
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Olga
authors.initials: O
authors.lastname: Levin-Kravets
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Ilan
authors.initials: I
authors.lastname: Attali
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3800, Australia.|Department of Biology, Technion-Israel Institute of Technology, Haifa 32000, Israel.
authors.forename: Oded
authors.initials: O
authors.lastname: Kleifeld
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Noa
authors.initials: N
authors.lastname: Shohat
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Shay
authors.initials: S
authors.lastname: Artzi
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Ori
authors.initials: O
authors.lastname: Zucker
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Inbar
authors.initials: I
authors.lastname: Pilzer
authors.affiliation: Department of Biology, Technion-Israel Institute of Technology, Haifa 32000, Israel.
authors.forename: Noa
authors.initials: N
authors.lastname: Reis
authors.affiliation: Department of Biology, Technion-Israel Institute of Technology, Haifa 32000, Israel.
authors.forename: Michael H
authors.initials: MH
authors.lastname: Glickman
authors.affiliation: Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel.
authors.forename: Shay
authors.initials: S
authors.lastname: Ben-Aroya
authors.affiliation: Department of Biochemistry and Molecular Biology and the Institute of Structural Biology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.|Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Gali
authors.identifier: 0000-0003-4703-7161
authors.initials: G
authors.lastname: Prag
mesh_terms: D004247:DNA / Q000032:analysis; D015966:Gene Expression Regulation, Fungal; D008854:Microscopy, Electron; D008968:Molecular Conformation; D016366:Open Reading Frames; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D000072417:Protein Domains; D025941:Protein Interaction Mapping; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D025801:Ubiquitin / Q000737:chemistry*; D054875:Ubiquitination
chemical_list: C122019:RPN10 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; D004247:DNA; D046988:Proteasome Endopeptidase Complex; C000597966:Rpn9 protein, S cerevisiae