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Canonical cytosolic iron-sulfur cluster assembly and non-canonical functions of DRE2 in Arabidopsis

机译:拟南芥中典型的胞质铁硫簇组装和DRE2的非典型功能

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摘要

As a component of the Cytosolic Iron-sulfur cluster Assembly (CIA) pathway, DRE2 is essential in organisms from yeast to mammals. However, the roles of DRE2 remain incompletely understood largely due to the lack of viable dre2 mutants. In this study, we successfully created hypomorphic dre2 mutants using the CRISPR/Cas9 technology. Like other CIA pathway mutants, the dre2 mutants have accumulation of DNA lesions and show constitutive DNA damage response. In addition, the dre2 mutants exhibit DNA hypermethylation at hundreds of loci. The mutant forms of DRE2 in the dre2 mutants, which bear deletions in the linker region of DRE2, lost interaction with GRXS17 but have stronger interaction with NBP35, resulting in the CIA-related defects of dre2. Interestingly, we find that DRE2 is also involved in auxin response that may be independent of its CIA role. DRE2 localizes in both the cytoplasm and the nucleus and nuclear DRE2 associates with euchromatin. Furthermore, DRE2 directly associates with multiple auxin responsive genes and maintains their normal expression. Our study highlights the importance of the linker region of DRE2 in coordinating CIA-related protein interactions and identifies the canonical and non-canonical roles of DRE2 in maintaining genome stability, epigenomic patterns, and auxin response.
机译:DRE2是胞质铁硫簇组装(CIA)途径的组成部分,在从酵母到哺乳动物的生物中都是必不可少的。但是,由于缺乏可行的dre2突变体,对DRE2的作用仍未完全了解。在这项研究中,我们使用CRISPR / Cas9技术成功创建了亚型dre2突变体。与其他CIA途径突变体一样,dre2突变体具有DNA损伤的积累,并显示出组成型DNA损伤反应。此外,dre2突变体在数百个位点显示DNA超甲基化。 dre2突变体中DRE2的突变体形式在DRE2的连接子区域带有缺失,与GRXS17失去相互作用,但与NBP35的相互作用更强,从而导致CIA相关的dre2缺陷。有趣的是,我们发现DRE2也参与了植物生长素的应答,这可能与其CIA的作用无关。 DRE2位于细胞质和细胞核中,核DRE2与常染色质相关。此外,DRE2与多个生长素反应基因直接缔合并维持其正常表达。我们的研究突出了DRE2接头区域在协调CIA相关蛋白相互作用中的重要性,并确定了DRE2在维持基因组稳定性,表观基因组模式和生长素反应中的规范作用和非规范作用。

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