Fluorescent Reporters for Ubiquitin-Dependent Proteolysis in Plants.
| Title: | Fluorescent Reporters for Ubiquitin-Dependent Proteolysis in Plants. |
|---|---|
| Authors: | Zientara-Rytter K; Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Pawinskiego 5A, 02-106, Warsaw, Poland.; Sirko A; Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Pawinskiego 5A, 02-106, Warsaw, Poland. asirko@ibb.waw.pl. |
| Source: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2016; Vol. 1450, pp. 45-54. |
| Publication Type: | Journal Article; Research Support, Non-U.S. Gov't |
| Language: | English |
| Journal Info: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| Imprint Name(s): | Publication: Totowa, NJ : Humana Press; Original Publication: Clifton, N.J. : Humana Press |
| MeSH Terms: | Proteolysis*; Molecular Biology/*methods ; Proteins/*chemistry ; Ubiquitin/*chemistry; Fluorescent Dyes/chemistry ; Plasmids/chemistry ; Proteasome Endopeptidase Complex/chemistry ; Proteins/isolation & purification ; Ubiquitin/biosynthesis ; Gene Expression Regulation, Plant |
| 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. |
| Contributed Indexing: | Keywords: Fluorescent reporters for ubiquitin; UFD; UPS |
| Substance Nomenclature: | 0 (Fluorescent Dyes); 0 (Proteins); 0 (Ubiquitin); EC 3.4.25.1 (Proteasome Endopeptidase Complex) |
| Entry Date(s): | Date Created: 20160719 Date Completed: 20171226 Latest Revision: 20180330 |
| Update Code: | 20260130 |
| DOI: | 10.1007/978-1-4939-3759-2_5 |
| PMID: | 27424745 |
| Database: | MEDLINE |
Journal Article; Research Support, Non-U.S. Gov't