title: Fluorescent Reporters for Ubiquitin-Dependent Proteolysis in Plants.
abstract: Ubiquitin is a small protein commonly used as a signal molecule which upon attachment to the proteins affects their function and their fate in the cells. For example, it can be used as a degradation marker by the cell. Ubiquitin plays a significant role in regulation of numerous cellular processes. Therefore, monitoring of ubiquitin-dependent proteolysis can provide important information. Here, we describe construction of YFP-based proteasome substrates containing modified ubiquitin and the protocol for their transient expression in plant cells for functional analysis of the ubiquitin/proteasome system. To facilitate further subcloning all plasmids generated by us are based on the Gateway(®) Cloning Technology and are compatible with the Gateway(®) destination vectors.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Pawinskiego 5A, 02-106, Warsaw, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Zientara-Rytter
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Pawinskiego 5A, 02-106, Warsaw, Poland. asirko@ibb.waw.pl.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Sirko
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry; D018506:Gene Expression Regulation, Plant; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000737:chemistry; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D059748:Proteolysis; D025801:Ubiquitin / Q000096:biosynthesis / Q000737:chemistry*
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex
keywords: Fluorescent reporters for ubiquitin; UFD; UPS