Conditional control of fluorescent protein degradation by an auxin-dependent nanobody.
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ICR Authors
Authors
Daniel, K
Icha, J
Horenburg, C
Müller, D
Norden, C
Mansfeld, J
Icha, J
Horenburg, C
Müller, D
Norden, C
Mansfeld, J
Document Type
Journal Article
Date
2018-08-17
Date Accepted
2018-07-27
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.
Citation
Nature communications, 2018, 9 (1), pp. 3297 - ?
Source Title
Publisher
NATURE PUBLISHING GROUP
ISSN
2041-1723
eISSN
2041-1723
Collections
Research Team
Post-translational modifications and cell proliferation