PUBLICATION
Conditional control of fluorescent protein degradation by an auxin-dependent nanobody
- Authors
- Daniel, K., Icha, J., Horenburg, C., Müller, D., Norden, C., Mansfeld, J.
- ID
- ZDB-PUB-180820-11
- Date
- 2018
- Source
- Nature communications 9: 3297 (Journal)
- Registered Authors
- Icha, Jaroslav, Norden, Caren
- Keywords
- none
- MeSH Terms
-
- Anaphase-Promoting Complex-Cyclosome/metabolism
- Animals
- Cell Compartmentation
- Green Fluorescent Proteins/metabolism*
- HeLa Cells
- Humans
- Indoleacetic Acids/metabolism*
- Kinetics
- Lysine/metabolism
- Proteolysis*
- Recombinant Fusion Proteins/metabolism
- Single-Domain Antibodies/metabolism*
- Zebrafish/metabolism
- PubMed
- 30120238 Full text @ Nat. Commun.
Citation
Daniel, K., Icha, J., Horenburg, C., Müller, D., Norden, C., Mansfeld, J. (2018) Conditional control of fluorescent protein degradation by an auxin-dependent nanobody. Nature communications. 9:3297.
Abstract
The conditional and reversible depletion of proteins by auxin-mediated degradation is a powerful tool to investigate protein functions in cells and whole organisms. However, its wider applications require fusing the auxin-inducible degron (AID) to individual target proteins. Thus, establishing the auxin system for multiple proteins can be challenging. Another approach for directed protein degradation are anti-GFP nanobodies, which can be applied to GFP stock collections that are readily available in different experimental models. Here, we combine the advantages of auxin and nanobody-based degradation technologies creating an AID-nanobody to degrade GFP-tagged proteins at different cellular structures in a conditional and reversible manner in human cells. We demonstrate efficient and reversible inactivation of the anaphase promoting complex/cyclosome (APC/C) and thus provide new means to study the functions of this essential ubiquitin E3 ligase. Further, we establish auxin degradation in a vertebrate model organism by employing AID-nanobodies in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping