A novel protein complex that regulates active DNA demethylation in Arabidopsis,Journal of Integrative Plant Biology

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A novel protein complex that regulates active DNA demethylation in Arabidopsis
Journal of Integrative Plant Biology ( IF 9.3 ) Pub Date : 2020-12-04 , DOI: 10.1111/jipb.13045
Pan Liu _{1,

2} , Wen-Feng Nie _{1,

3} , Xiansong Xiong _{1,

2} , Yuhua Wang ₁ , Yuwei Jiang _{1,

2} , Pei Huang _{1,

2} , Xueqiang Lin ₁ , Guochen Qin ₁ , Huan Huang ₁ , Qingfeng Niu ₁ , Jiamu Du ₁ , Zhaobo Lang ₁ , Rosa Lozano-Duran ₁ , Jian-Kang Zhu _{1,

4}

Affiliation

Active DNA demethylation is critical for altering DNA methylation patterns and regulating gene expression. The 5‐methylcytosine DNA glycosylase/lyase ROS1 initiates a base‐excision repair pathway for active DNA demethylation and is required for the prevention of DNA hypermethylation at 1 000s of genomic regions in Arabidopsis. How ROS1 is regulated and targeted to specific genomic regions is not well understood. Here, we report the discovery of an Arabidopsis protein complex that contains ROS1, regulates ROS1 gene expression, and likely targets the ROS1 protein to specific genomic regions. ROS1 physically interacts with a WD40 domain protein (RWD40), which in turn interacts with a methyl‐DNA binding protein (RMB1) as well as with a zinc finger and homeobox domain protein (RHD1). RMB1 binds to DNA that is methylated in any sequence context, and this binding is necessary for its function in vivo. Loss‐of‐function mutations in RWD40, RMB1, or RHD1 cause DNA hypermethylation at several tested genomic regions independently of the known ROS1 regulator IDM1. Because the hypermethylated genomic regions include the DNA methylation monitoring sequence in the ROS1 promoter, plants mutated in RWD40, RMB1, or RHD1 show increased ROS1 expression. Importantly, ROS1 binding to the ROS1 promoter requires RWD40, RMB1, and RHD1, suggesting that this complex dictates ROS1 targeting to this locus. Our results demonstrate that ROS1 forms a protein complex with RWD40, RMB1, and RHD1, and that this novel complex regulates active DNA demethylation at several endogenous loci in Arabidopsis.

中文翻译：

一种调节拟南芥活性 DNA 去甲基化的新型蛋白质复合物

主动 DNA 去甲基化对于改变 DNA 甲基化模式和调节基因表达至关重要。5-甲基胞嘧啶 DNA 糖基化酶/裂解酶 ROS1 启动碱基切除修复途径以进行活性 DNA 去甲基化，并且是预防拟南芥基因组区域 1000 处 DNA 高甲基化所必需的。ROS1 是如何被调节和靶向特定基因组区域的，目前还不清楚。在这里，我们报告了一种含有 ROS1的拟南芥蛋白复合物的发现，调节ROS1基因表达，并且可能将 ROS1 蛋白靶向特定的基因组区域。ROS1 与 WD40 结构域蛋白 (RWD40) 发生物理相互作用，后者又与甲基 DNA 结合蛋白 (RMB1) 以及锌指和同源框结构域蛋白 (RHD1) 相互作用。RMB1 与在任何序列环境中甲基化的 DNA 结合，这种结合对于其体内功能是必需的。RWD40、RMB1或RHD1中的功能丧失突变导致几个测试基因组区域的 DNA 高甲基化，独立于已知的 ROS1 调节因子 IDM1。由于高甲基化基因组区域包括ROS1启动子中的 DNA 甲基化监测序列，因此在RWD40 中发生突变的植物、RMB1或RHD1显示ROS1表达增加。重要的是，ROS1 与ROS1启动子的结合需要 RWD40、RMB1 和 RHD1，这表明该复合物决定了 ROS1 靶向该基因座。我们的结果表明 ROS1 与 RWD40、RMB1 和 RHD1 形成蛋白质复合物，并且这种新型复合物调节拟南芥中几个内源性位点的活性 DNA 去甲基化。

更新日期：2020-12-04

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