Rice ( Oryza sativa ) TIR1 and 5'adamantyl-IAA Significantly Improve the Auxin-Inducible Degron System in  Schizosaccharomyces pombe .
The 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 F74A , 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.
Watson AT, Hassell-Hart S, Spencer J, Carr AM