title: Dissecting ubiquitin signaling with linkage-defined and protease resistant ubiquitin chains.
abstract: Ubiquitylation is a complex posttranslational protein modification and deregulation of this pathway has been associated with different human disorders. Ubiquitylation comes in different flavors: Besides mono-ubiquitylation, ubiquitin chains of various topologies are formed on substrate proteins. The fate of ubiquitylated proteins is determined by the linkage-type of the attached ubiquitin chains, however, the underlying mechanism is poorly characterized. Herein, we describe a new method based on codon expansion and click-chemistry-based polymerization to generate linkage-defined ubiquitin chains that are resistant to ubiquitin-specific proteases and adopt native-like functions. The potential of these artificial chains for analyzing ubiquitin signaling is demonstrated by linkage-specific effects on cell-cycle progression.
authors.affiliation: Departments of Chemistry and Biology, Konstanz Research School Chemical Biology, University of Konstanz, Universitätsstrasse 10, 78457 Konstanz (Germany).
authors.forename: Tatjana
authors.initials: T
authors.lastname: Schneider
authors.forename: Daniel
authors.initials: D
authors.lastname: Schneider
authors.forename: Daniel
authors.initials: D
authors.lastname: Rösner
authors.forename: Saurav
authors.initials: S
authors.lastname: Malhotra
authors.forename: Franziska
authors.initials: F
authors.lastname: Mortensen
authors.forename: Thomas U
authors.initials: TU
authors.lastname: Mayer
authors.forename: Martin
authors.initials: M
authors.lastname: Scheffner
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
mesh_terms: D000818:Animals; D057930:Click Chemistry; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D058105:Polymerization; D015398:Signal Transduction; D025801:Ubiquitin / Q000737:chemistry* / Q000378:metabolism*; D054875:Ubiquitination; D014981:Xenopus
chemical_list: D025801:Ubiquitin; D010447:Peptide Hydrolases
keywords: click chemistry; codon expansion; posttranslational modification; ubiquitin chains; unnatural amino acids