m⁶A modification of mutant huntingtin RNA promotes the biogenesis of pathogenic huntingtin transcripts.

IF 6.5 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY EMBO Reports Pub Date : 2024-11-01 Epub Date: 2024-10-11 DOI:10.1038/s44319-024-00283-7

Anika Pupak, Irene Rodríguez-Navarro, Kirupa Sathasivam, Ankita Singh, Amelie Essmann, Daniel Del Toro, Silvia Ginés, Ricardo Mouro Pinto, Gillian P Bates, Ulf Andersson Vang Ørom, Eulàlia Martí, Verónica Brito

{"title":"m6A modification of mutant huntingtin RNA promotes the biogenesis of pathogenic huntingtin transcripts.","authors":"Anika Pupak, Irene Rodríguez-Navarro, Kirupa Sathasivam, Ankita Singh, Amelie Essmann, Daniel Del Toro, Silvia Ginés, Ricardo Mouro Pinto, Gillian P Bates, Ulf Andersson Vang Ørom, Eulàlia Martí, Verónica Brito","doi":"10.1038/s44319-024-00283-7","DOIUrl":null,"url":null,"abstract":"In Huntington's disease (HD), aberrant processing of huntingtin (HTT) mRNA produces HTT1a transcripts that encode the pathogenic HTT exon 1 protein. The mechanisms behind HTT1a production are not fully understood. Considering the role of m6A in RNA processing and splicing, we investigated its involvement in HTT1a generation. Here, we show that m6A methylation is increased before the cryptic poly(A) sites (IpA1 and IpA2) within the huntingtin RNA in the striatum of Hdh+/Q111 mice and human HD samples. We further assessed m6A's role in mutant Htt mRNA processing by pharmacological inhibition and knockdown of METTL3, as well as targeted demethylation of Htt intron 1 using a dCas13-ALKBH5 system in HD mouse cells. Our data reveal that Htt1a transcript levels are regulated by both METTL3 and the methylation status of Htt intron 1. They also show that m6A methylation in intron 1 depends on expanded CAG repeats. Our findings highlight a potential role for m6A in aberrant splicing of Htt mRNA.","PeriodicalId":11541,"journal":{"name":"EMBO Reports","volume":" ","pages":"5026-5052"},"PeriodicalIF":6.5000,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11549361/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"EMBO Reports","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1038/s44319-024-00283-7","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/10/11 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

In Huntington's disease (HD), aberrant processing of huntingtin (HTT) mRNA produces HTT1a transcripts that encode the pathogenic HTT exon 1 protein. The mechanisms behind HTT1a production are not fully understood. Considering the role of m⁶A in RNA processing and splicing, we investigated its involvement in HTT1a generation. Here, we show that m⁶A methylation is increased before the cryptic poly(A) sites (IpA1 and IpA2) within the huntingtin RNA in the striatum of Hdh+/Q111 mice and human HD samples. We further assessed m⁶A's role in mutant Htt mRNA processing by pharmacological inhibition and knockdown of METTL3, as well as targeted demethylation of Htt intron 1 using a dCas13-ALKBH5 system in HD mouse cells. Our data reveal that Htt1a transcript levels are regulated by both METTL3 and the methylation status of Htt intron 1. They also show that m⁶A methylation in intron 1 depends on expanded CAG repeats. Our findings highlight a potential role for m⁶A in aberrant splicing of Htt mRNA.

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

突变亨廷蛋白 RNA 的 m6A 修饰促进了致病亨廷蛋白转录本的生物生成。

在亨廷顿氏病（HD）中，亨廷汀（HTT）mRNA 的异常加工会产生 HTT1a 转录本，编码致病的 HTT 第 1 外显子蛋白。HTT1a产生的机制尚未完全明了。考虑到 m6A 在 RNA 处理和剪接中的作用，我们研究了它在 HTT1a 生成中的参与。在这里，我们发现在 Hdh+/Q111 小鼠和人类 HD 样本的纹状体中，m6A 甲基化在亨廷蛋白 RNA 中的隐性 poly(A) 位点（IpA1 和 IpA2）之前增加。我们进一步评估了 m6A 在突变型 Htt mRNA 处理中的作用，方法是药理抑制和敲除 METTL3，以及使用 dCas13-ALKBH5 系统在 HD 小鼠细胞中对 Htt 内含子 1 进行靶向去甲基化。我们的数据显示，Htt1a 转录水平受 METTL3 和 Htt 内含子 1 甲基化状态的调控。这些数据还显示，内含子 1 中的 m6A 甲基化取决于扩展的 CAG 重复序列。我们的发现突显了 m6A 在 Htt mRNA 的异常剪接中的潜在作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

EMBO Reports 生物-生化与分子生物学

CiteScore

11.20

自引率

1.30%

发文量

267

审稿时长

1 months

期刊介绍： EMBO Reports is a scientific journal that specializes in publishing research articles in the fields of molecular biology, cell biology, and developmental biology. The journal is known for its commitment to publishing high-quality, impactful research that provides novel physiological and functional insights. These insights are expected to be supported by robust evidence, with independent lines of inquiry validating the findings. The journal's scope includes both long and short-format papers, catering to different types of research contributions. It values studies that: Communicate major findings: Articles that report significant discoveries or advancements in the understanding of biological processes at the molecular, cellular, and developmental levels. Confirm important findings: Research that validates or supports existing knowledge in the field, reinforcing the reliability of previous studies. Refute prominent claims: Studies that challenge or disprove widely accepted ideas or hypotheses in the biosciences, contributing to the correction and evolution of scientific understanding. Present null data: Papers that report negative results or findings that do not support a particular hypothesis, which are crucial for the scientific process as they help to refine or redirect research efforts. EMBO Reports is dedicated to maintaining high standards of scientific rigor and integrity, ensuring that the research it publishes contributes meaningfully to the advancement of knowledge in the life sciences. By covering a broad spectrum of topics and encouraging the publication of both positive and negative results, the journal plays a vital role in promoting a comprehensive and balanced view of scientific inquiry.