The RNA-binding domain of DCL3 is required for long-distance RNAi signaling

IF 4.6 4区 农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Jie Li, Bo-Sen Zhang, Hua-Wei Wu, Cheng-Lan Liu, Hui-Shan Guo, Jian-Hua Zhao
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引用次数: 0

Abstract

Small RNA (sRNA)-mediated RNA silencing (also known as RNA interference, or RNAi) is a conserved mechanism in eukaryotes that includes RNA degradation, DNA methylation, heterochromatin formation and protein translation repression. In plants, sRNAs can move either cell-to-cell or systemically, thereby acting as mobile silencing signals to trigger noncell autonomous silencing. However, whether and what proteins are also involved in noncell autonomous silencing have not been elucidated. In this study, we utilized a previously reported inducible RNAi plant, PDSi, which can induce systemic silencing of the endogenous PDS gene, and we demonstrated that DCL3 is involved in systemic PDS silencing through its RNA binding activity. We confirmed that the C-terminus of DCL3, including the predicted RNA-binding domain, is capable of binding short RNAs. Mutations affecting RNA binding, but not processing activity, reduced systemic PDS silencing, indicating that DCL3 binding to RNAs is required for the induction of systemic silencing. Cucumber mosaic virus infection assays showed that the RNA-binding activity of DCL3 is required for antiviral RNAi in systemically noninoculated leaves. Our findings demonstrate that DCL3 acts as a signaling agent involved in noncell autonomous silencing and an antiviral effect in addition to its previously known function in the generation of 24-nucleotide sRNAs.

DCL3的RNA结合域是长距离RNAi信号传导所必需的。
小 RNA(sRNA)介导的 RNA 沉默(也称为 RNA 干扰或 RNAi)是真核生物中的一种保守机制,包括 RNA 降解、DNA 甲基化、异染色质形成和蛋白质翻译抑制。在植物中,sRNA 可以在细胞间移动或系统移动,从而作为移动沉默信号触发非细胞自主沉默。然而,非细胞自主沉默中是否也有蛋白质参与以及参与的蛋白质是什么,这些问题尚未得到阐明。在本研究中,我们利用之前报道的可诱导的 RNAi 植物 PDSi,它可以诱导内源 PDS 基因的系统沉默,我们证明了 DCL3 通过其 RNA 结合活性参与了 PDS 的系统沉默。我们证实 DCL3 的 C 端(包括预测的 RNA 结合域)能够结合短 RNA。影响 RNA 结合而非处理活性的突变降低了系统性 PDS 沉默,这表明 DCL3 与 RNA 的结合是诱导系统性沉默所必需的。黄瓜花叶病毒感染试验表明,DCL3的RNA结合活性是系统非接种叶片抗病毒RNAi所必需的。我们的研究结果表明,除了之前已知的生成 24 核苷酸 sRNA 的功能外,DCL3 还是一种参与非细胞自主沉默和抗病毒作用的信号传导因子:在线版本包含补充材料,见 10.1007/s42994-023-00124-6。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
7.70
自引率
2.80%
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0
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