（）TIR1 和 5'adamantyl-IAA 显著提高了 在 中的生长素诱导型降解结构域系统。
Rice () TIR1 and 5'adamantyl-IAA Significantly Improve the Auxin-Inducible Degron System in .
机构信息
Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Brighton BN1 9RQ, UK.
Department of Chemistry, School of Life Sciences, University of Sussex, Brighton BN1 9QJ, UK.
出版信息
Genes (Basel). 2021 Jun 8;12(6):882. doi: 10.3390/genes12060882.
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 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 that uses the TIR1 from (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.
生长素诱导的降解结构域（AID）系统是一种在真核模式生物中诱导靶向蛋白降解的强大工具。由于 F-box 蛋白 TIR1 蛋白与 SCF 成分 Skp1（Skp1-TIR1）融合，现有的 AID 系统效率有限。在这里，我们报告了一种改进的 AID 系统，该系统使用来自 （OsTIR1）的 TIR1 而不与 Skp1 融合。此外，我们证明通过将 OsTIR1 生长素结合位点突变体 OsTIR1 与生长素类似物 5' -adamantyl-IAA（AID2）配对，可以提高降解效率。我们通过将必需的 DNA 复制因子 Mcm4 和非必需的重组蛋白 Rad52 应用于 OsTIR1 AID 和 AID2 系统，提供了增强的功能证据。与 AID 不同，使用 AID2 时未观察到可检测到的生长素非依赖性 AID 标记蛋白耗竭。
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