genes-12-00882.pdf (1.82 MB)
Rice (Oryza sativa) TIR1 and 5'adamantyl-IAA significantly improve the auxin-inducible degron system in Schizosaccharomyces pombe
journal contribution
posted on 2023-06-10, 00:03 authored by Adam WatsonAdam Watson, Storm Hassell-HartStorm Hassell-Hart, John SpencerJohn Spencer, Antony CarrAntony CarrThe auxin-inducible degron (AID) system is a powerful tool to induce targeted degradation of proteins in eukaryotic model organisms. The efficiency of the existing Schizosaccharomyces pombe AID system is limited due to the fusion of the F-box protein TIR1 protein to the SCF component, Skp1 (Skp1-TIR1). Here, we report an improved AID system for S. pombe that uses the TIR1 from Oryza sativa (OsTIR1) not fused to Skp1. Furthermore, we demonstrate that degradation efficiency can be improved by pairing an OsTIR1 auxin-binding site mutant, OsTIR1, with an auxin analogue, 5'adamantyl-IAA (AID2). We provide evidence for the enhanced functionality of the OsTIR1 AID and AID2 systems by application to the essential DNA replication factor Mcm4 and to a non-essential recombination protein, Rad52. Unlike AID, no detectable auxin-independent depletion of AID-tagged proteins was observed using AID2
Funding
Poised Fragment Libraries for Atypical Bromodomain Inhibition; G2210; EPSRC-ENGINEERING & PHYSICAL SCIENCES RESEARCH COUNCIL; EP/P026990/1
History
Publication status
- Published
File Version
- Published version
Journal
GenesISSN
2073-4425Publisher
MDPIExternal DOI
Issue
6Volume
12Page range
1-23Article number
a882Pages
23.0Department affiliated with
- Chemistry Publications
Full text available
- Yes
Peer reviewed?
- Yes
Legacy Posted Date
2021-06-09First Open Access (FOA) Date
2021-06-09First Compliant Deposit (FCD) Date
2021-06-08Usage metrics
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