DTF1 is a core component of RNA-directed DNA methylation and may assist in the recruitment of Pol IV

  • Heng Zhang
    Shanghai Center for Plant Stress Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;
  • Ze-Yang Ma
    National Institute of Biological Sciences, Beijing 102206, China;
  • Liang Zeng
    Shanghai Center for Plant Stress Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;
  • Kaori Tanaka
    Department of Biochemistry and Molecular Biology, Division of Basic Science Research, University of Texas MD Anderson Cancer Center, Houston, TX 77030;
  • Cui-Jun Zhang
    National Institute of Biological Sciences, Beijing 102206, China;
  • Jun Ma
    Shanghai Center for Plant Stress Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;
  • Ge Bai
    Shanghai Center for Plant Stress Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;
  • Pengcheng Wang
    Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907; and
  • Su-Wei Zhang
    National Institute of Biological Sciences, Beijing 102206, China;
  • Zhang-Wei Liu
    National Institute of Biological Sciences, Beijing 102206, China;
  • Tao Cai
    National Institute of Biological Sciences, Beijing 102206, China;
  • Kai Tang
    Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907; and
  • Renyi Liu
    Department of Botany and Plant Sciences, University of California, Riverside, CA 92521
  • Xiaobing Shi
    Department of Biochemistry and Molecular Biology, Division of Basic Science Research, University of Texas MD Anderson Cancer Center, Houston, TX 77030;
  • Xin-Jian He
    National Institute of Biological Sciences, Beijing 102206, China;
  • Jian-Kang Zhu
    Shanghai Center for Plant Stress Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China;

抄録

<jats:p> DNA methylation is an important epigenetic mark in many eukaryotic organisms. De novo DNA methylation in plants can be achieved by the RNA-directed DNA methylation (RdDM) pathway, where the plant-specific DNA-dependent RNA polymerase IV (Pol IV) transcribes target sequences to initiate 24-nt siRNA production and action. The putative DNA binding protein DTF1/SHH1 of <jats:italic>Arabidopsis</jats:italic> has been shown to associate with Pol IV and is required for 24-nt siRNA accumulation and transcriptional silencing at several RdDM target loci. However, the extent and mechanism of DTF1 function in RdDM is unclear. We show here that DTF1 is necessary for the accumulation of the majority of Pol IV-dependent 24-nt siRNAs. It is also required for a large proportion of Pol IV-dependent de novo DNA methylation. Interestingly, there is a group of RdDM target loci where 24-nt siRNA accumulation but not DNA methylation is dependent on DTF1. DTF1 interacts directly with the chromatin remodeling protein CLASSY 1 (CLSY1), and both DTF1 and CLSY1 are associated in vivo with Pol IV but not Pol V, which functions downstream in the RdDM effector complex. DTF1 and DTF2 (a DTF1-like protein) contain a SAWADEE domain, which was found to bind specifically to histone H3 containing H3K9 methylation. Taken together, our results show that DTF1 is a core component of the RdDM pathway, and suggest that DTF1 interacts with CLSY1 to assist in the recruitment of Pol IV to RdDM target loci where H3K9 methylation may be an important feature. Our results also suggest the involvement of DTF1 in an important negative feedback mechanism for DNA methylation at some RdDM target loci. </jats:p>

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