| Chapter title |
Proteostasis
|
|---|---|
| Chapter number | 12 |
| Book title |
Proteostasis
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3756-1_12 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3754-7, 978-1-4939-3756-1
|
| Authors |
Matilainen, Olli, Jha, Sweta, Holmberg, Carina I, Olli Matilainen, Sweta Jha, Carina I. Holmberg, Holmberg, Carina I. |
| Editors |
Rune Matthiesen |
| Abstract |
The ubiquitin-proteasome system (UPS) plays a key role in maintaining proteostasis by degrading most of the cellular proteins. Traditionally, UPS activity is studied in vitro, in yeast, or in mammalian cell cultures by using short-lived GFP-based UPS reporters. Here, we present protocols for two fluorescent tools facilitating real-time imaging of UPS activity in living animals. We have generated transgenic Caenorhabditis elegans (C. elegans) expressing a photoconvertible UbG76V-Dendra2 UPS reporter, which permits measurement of reporter degradation by the proteasome independently of reporter protein synthesis, and a fluorescent polyubiquitin-binding reporter for detection of the endogenous pool of Lys48-linked polyubiquitinated proteasomal substrates. These reporter systems facilitate cell- and tissue-specific analysis of UPS activity especially in young adult animals, but can also be used for studies during development, aging, and for example stress conditions. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 1 | 20% |
| Germany | 1 | 20% |
| Unknown | 3 | 60% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 5 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 18 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 5 | 28% |
| Student > Ph. D. Student | 3 | 17% |
| Professor > Associate Professor | 2 | 11% |
| Student > Doctoral Student | 1 | 6% |
| Lecturer > Senior Lecturer | 1 | 6% |
| Other | 3 | 17% |
| Unknown | 3 | 17% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 7 | 39% |
| Agricultural and Biological Sciences | 5 | 28% |
| Medicine and Dentistry | 2 | 11% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 6% |
| Unknown | 3 | 17% |