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

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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

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

Genes

ISSN

2073-4425

Publisher

MDPI

Issue

6

Volume

12

Page range

1-23

Article number

a882

Pages

23.0

Department affiliated with

  • Chemistry Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2021-06-09

First Open Access (FOA) Date

2021-06-09

First Compliant Deposit (FCD) Date

2021-06-08

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