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

Canonical cytosolic iron-sulfur cluster assembly and non-canonical functions of DRE2 in Arabidopsis
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拟南芥中 DRE2 的典型胞质铁硫簇组装和非典型功能

DOI:
10.1371/journal.pgen.1008094
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发表时间:
2019-04
期刊:
影响因子:
4.5
通讯作者:
Qian Weiqiang
Qian Weiqiang
中科院分区:
生物学2区
文献类型:
--
作者:
Wang Xiaokang;Chen Xudong;Sun Linhua;Qian Weiqiang

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DRE2作为胞质铁硫簇组装(CIA)途径的一个组成部分,在从酵母到哺乳动物的生物体中都是必不可少的。然而,由于缺乏可行的DRE2突变体,DRE2的作用仍然不完全清楚。在本研究中,我们利用CRISPR/Cas9技术成功构建了半胚型的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,并表现出本构性DNA损伤反应。此外,dre2突变体在数百个位点表现出DNA超甲基化。在DRE2突变体中,DRE2的突变体形式在DRE2的连接子区域有缺失,与GRXS17失去相互作用,但与NBP35有更强的相互作用,导致DRE2的cia相关缺陷。有趣的是,我们发现DRE2也参与生长素反应,可能独立于CIA的作用。DRE2定位于细胞质和细胞核中,细胞核中DRE2与常染色质相关。此外,DRE2直接与多种生长素应答基因结合,维持其正常表达。我们的研究强调了DRE2连接区域在协调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典型和非典型作用。
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.
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