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研究通过被子植物和苔藓植物 HEN1 的功能比较来抑制小 RNA 甲基化的病毒抑制剂 HC-Pro。

Investigating the Viral Suppressor HC-Pro Inhibiting Small RNA Methylation through Functional Comparison of HEN1 in Angiosperm and Bryophyte.

机构信息

Institute of Biotechnology, National Taiwan University, Taipei 106, Taiwan.

The PhD Program for Translational Medicine, College of Medical Science and Technology, Taipei Medical University and Academia Sinica, Taipei 115, Taiwan.

出版信息

Viruses. 2021 Sep 15;13(9):1837. doi: 10.3390/v13091837.

DOI:10.3390/v13091837
PMID:34578418
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8473176/
Abstract

In plants, HEN1-facilitated methylation at 3' end ribose is a critical step of small-RNA (sRNA) biogenesis. A mutant of well-studied Arabidopsis HEN1 (AtHEN1), , showed a defective developmental phenotype, indicating the importance of sRNA methylation. Moreover, has been identified to have a ortholog gene (Mp); however, its function remained unfathomed. Our in vivo and in vitro data have shown MpHEN1 activity being comparable with AtHEN1, and their substrate specificity towards duplex microRNA (miRNA) remained consistent. Furthermore, the phylogenetic tree and multiple alignment highlighted the conserved molecular evolution of the HEN1 family in plants. The P1/HC-Pro of the turnip mosaic virus (TuMV) is a known RNA silencing suppressor and inhibits HEN1 methylation of sRNAs. Here, we report that the HC-Pro physically binds with AtHEN1 through FRNK motif, inhibiting HEN1's methylation activity. Moreover, the in vitro EMSA data indicates GST-HC-Pro of TuMV lacks sRNA duplex-binding ability. Surprisingly, the HC-Pro also inhibits MpHEN1 activity in a dosage-dependent manner, suggesting the possibility of interaction between HC-Pro and MpHEN1 as well. Further investigations on understanding interaction mechanisms of HEN1 and various HC-Pros can advance the knowledge of viral suppressors.

摘要

在植物中，HEN1 介导的 3' 端核糖甲基化是小 RNA（sRNA）生物发生的关键步骤。一个经过充分研究的拟南芥 HEN1（AtHEN1）突变体，表现出发育缺陷表型，表明 sRNA 甲基化的重要性。此外，已经鉴定出具有一个 同源基因（Mp）；然而，其功能仍未被揭示。我们的体内和体外数据表明 MpHEN1 活性与 AtHEN1 相当，并且它们对双链 microRNA（miRNA）的底物特异性保持一致。此外，系统发育树和多重比对突出了植物 HEN1 家族的保守分子进化。芜菁花叶病毒（TuMV）的 P1/HC-Pro 是一种已知的 RNA 沉默抑制剂，抑制 sRNA 的 HEN1 甲基化。在这里，我们报告 TuMV 的 HC-Pro 通过 FRNK 基序与 AtHEN1 物理结合，抑制 HEN1 的甲基化活性。此外，体外 EMSA 数据表明 TuMV 的 GST-HC-Pro 缺乏 sRNA 双链结合能力。令人惊讶的是，HC-Pro 也以剂量依赖的方式抑制 MpHEN1 活性，表明 HC-Pro 与 MpHEN1 之间也可能存在相互作用。进一步研究 HEN1 和各种 HC-Pro 之间的相互作用机制可以提高对病毒抑制剂的认识。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/98472f1f12be/viruses-13-01837-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/647bae8a7577/viruses-13-01837-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/61491d26c438/viruses-13-01837-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/f4e388ff214a/viruses-13-01837-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/6180878a5bf7/viruses-13-01837-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/e600cc96ca65/viruses-13-01837-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/88684d33827c/viruses-13-01837-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/25bccfd1f8e0/viruses-13-01837-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/902e07651ee1/viruses-13-01837-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/5b76ea5a7df8/viruses-13-01837-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/98472f1f12be/viruses-13-01837-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/647bae8a7577/viruses-13-01837-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/61491d26c438/viruses-13-01837-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/f4e388ff214a/viruses-13-01837-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/6180878a5bf7/viruses-13-01837-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/e600cc96ca65/viruses-13-01837-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/88684d33827c/viruses-13-01837-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/25bccfd1f8e0/viruses-13-01837-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/902e07651ee1/viruses-13-01837-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/5b76ea5a7df8/viruses-13-01837-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cde1/8473176/98472f1f12be/viruses-13-01837-g010.jpg

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