University School of Biotechnology, Guru Gobind Singh Indraprastha University, New Delhi, India.
Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, India.
FEBS J. 2024 Jan;291(1):92-113. doi: 10.1111/febs.16935. Epub 2023 Aug 29.
TRDMT1/DNMT2 belongs to the conserved family of nucleic acid methyltransferases. Unlike the animal systems, studies on TRDMT1/DNMT2 in land plants have been limited. We show that TRDMT1/DNMT2 is strongly conserved in the green lineage. Studies in mosses have previously shown that TRDMT1/DNMT2 plays a crucial role in modulating molecular networks involved in stress perception and signalling and in transcription/stability of specific tRNAs under stress. To gain deeper insight into its biological roles in a flowering plant, we examined more closely the previously reported Arabidopsis SALK_136635C line deficient in TRDMT1/DNMT2 function [Goll MG et al. (2006) Science 311, 395-398]. RNAs derived from Arabidopsis Dnmt2-deficient plants lacked m C38 in tRNA . In this study, by transient expression assays we show that Arabidopsis TRDMT1/DNMT2 is distributed in the nucleus, cytoplasm and RNA-processing bodies, suggesting a role for TRDMT1/DNMT2 in RNA metabolic processes possibly by shuttling between cellular compartments. Bright-field and high-resolution SEM and qPCR analysis reveal roles of TRDMT1/DNMT2 in proper growth and developmental progression. Quantitative proteome analysis by LC-MS/MS coupled with qPCR shows AtTRDMT1/AtDNMT2 function to be crucial for protein synthesis and cellular homeostasis via housekeeping roles and proteins with poly-Asp stretches and RNA pol II activity on selected genes are affected in attrdmt1/atdnmt2. This shift in metabolic pathways primes the mutant plants to become increasingly sensitive to oxidative and osmotic stress. Taken together, our study sheds light on the mechanistic role of TRDMT1/DNMT2 in a flowering plant.
TRDMT1/DNMT2 属于保守的核酸甲基转移酶家族。与动物系统不同,对陆地植物中的 TRDMT1/DNMT2 的研究一直很有限。我们表明,TRDMT1/DNMT2 在绿色谱系中高度保守。先前在苔藓中的研究表明,TRDMT1/DNMT2 在调节涉及应激感知和信号转导的分子网络以及在应激下特定 tRNA 的转录/稳定性方面发挥着关键作用。为了更深入地了解其在开花植物中的生物学作用,我们更仔细地研究了先前报道的拟南芥 SALK_136635C 系,该系缺乏 TRDMT1/DNMT2 功能[Goll MG 等人。(2006 年)科学 311, 395-398]。来自拟南芥 Dnmt2 缺陷型植物的 RNA 缺乏 tRNA 中的 m C38。在这项研究中,通过瞬时表达测定,我们表明拟南芥 TRDMT1/DNMT2 分布在细胞核、细胞质和 RNA 处理体中,这表明 TRDMT1/DNMT2 可能在细胞区室之间穿梭,参与 RNA 代谢过程。明场和高分辨率 SEM 和 qPCR 分析揭示了 TRDMT1/DNMT2 在适当生长和发育进展中的作用。通过 LC-MS/MS 与 qPCR 结合的定量蛋白质组分析表明,AtTRDMT1/AtDNMT2 的功能对于通过管家作用和具有多-Asp 延伸的蛋白质以及选定基因上的 RNA pol II 活性对于蛋白质合成和细胞内稳态至关重要在 attrdmt1/atdnmt2 中受到影响。这种代谢途径的转变使突变体植物对氧化和渗透胁迫变得越来越敏感。总之,我们的研究揭示了 TRDMT1/DNMT2 在开花植物中的机制作用。