Rice (Oryza sativa) TIR1 and 5′adamantyl-IAA significantly improve the auxin-inducible degron system in Schizosaccharomyces pombe

Watson, Adam T, Hassell-Hart, Storm, Spencer, John and Carr, Antony M (2021) Rice (Oryza sativa) TIR1 and 5′adamantyl-IAA significantly improve the auxin-inducible degron system in Schizosaccharomyces pombe. Genes, 12 (6). a882 1-23. ISSN 2073-4425

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Abstract

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, 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

Item Type: Article
Schools and Departments: School of Life Sciences > Chemistry
SWORD Depositor: Mx Elements Account
Depositing User: Mx Elements Account
Date Deposited: 09 Jun 2021 16:00
Last Modified: 28 Feb 2022 16:18
URI: http://sro.sussex.ac.uk/id/eprint/99705

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Project NameSussex Project NumberFunderFunder Ref
Poised Fragment Libraries for Atypical Bromodomain InhibitionG2210EPSRC-ENGINEERING & PHYSICAL SCIENCES RESEARCH COUNCILEP/P026990/1