The importance of m(6)A topology in chicken embryo mRNA: a precise mapping of m(6)A at the conserved chicken β-actin zipcode.
The importance of m(6)A topology in chicken embryo mRNA: a precise mapping of m(6)A at the conserved chicken β-actin zipcode.
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DOI：
10.1261/rna.079615.123
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发表时间：
2023-06
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影响因子：
--
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mRNA中的N6-甲基腺苷（m6 A）几乎调节mRNA生命周期的每个阶段，使用m6 A特异性甲基化RNA免疫沉淀与下一代测序（MeRIPSeq）或m6 A单个核苷酸分辨率交联和免疫沉淀（miCLIP）高通量检测mRNA中甲基化位点的方法学的发展彻底改变了m6 A研究领域。这两种方法都是基于片段化mRNA的免疫沉淀。然而，有充分证据表明抗体通常具有非特异性活性，因此使用抗体非依赖性方法验证鉴定的m6 A位点将是高度期望的。我们基于来自鸡胚胎MeRIPSeq结果的数据和我们的RNA-表观修饰检测和碱基识别（RedBaron）抗体非依赖性测定，对鸡β-肌动蛋白邮政编码中的m6 A位点进行了定位和定量。我们还证明了β-肌动蛋白邮政编码中该位点的甲基化增强了ZBP 1的体外结合，而附近腺苷的甲基化则消除了结合。这表明m6 A可能在调节β-肌动蛋白mRNA的局部翻译中起作用，并且m6 A增强或抑制阅读器蛋白的RNA结合的能力突出了在核苷酸分辨率下检测m6 A的重要性。
N6-methyladenosine (m6A) in mRNA regulates almost every stage in the mRNA life cycle, and the development of methodologies for the high-throughput detection of methylated sites in mRNA using m6A-specific methylated RNA immunoprecipitation with next-generation sequencing (MeRIPSeq) or m6A individual-nucleotide-resolution cross-linking and immunoprecipitation (miCLIP) have revolutionized the m6A research field. Both of these methods are based on immunoprecipitation of fragmented mRNA. However, it is well documented that antibodies often have nonspecific activities, thus verification of identified m6A sites using an antibody-independent method would be highly desirable. We mapped and quantified the m6A site in the chicken β-actin zipcode based on the data from chicken embryo MeRIPSeq results and our RNA-Epimodification Detection and Base-Recognition (RedBaron) antibody-independent assay. We also demonstrated that methylation of this site in the β-actin zipcode enhances ZBP1 binding in vitro, while methylation of a nearby adenosine abolishes binding. This suggests that m6A may play a role in regulating localized translation of β-actin mRNA, and the ability of m6A to enhance or inhibit a reader protein's RNA binding highlights the importance of m6A detection at nucleotide resolution.
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DOI：
10.1261/rna.058206.116
发表时间：
2017-10
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Antanaviciute A;Baquero-Perez B;Watson CM;Harrison SM;Lascelles C;Crinnion L;Markham AF;Bonthron DT;Whitehouse A;Carr IM
通讯作者：
Carr IM
YTHDF2 通过直接招募 CCR4-NOT 去腺苷酶复合物来破坏含有 m(6)A 的 RNA 的稳定性。
影响因子：
16.6
作者：
Du, Hao;Zhao, Ya;He, Jinqiu;Zhang, Yao;Xi, Hairui;Liu, Mofang;Ma, Jinbiao;Wu, Ligang
通讯作者：
Wu, Ligang
影响因子：
48
作者：
Linder, Bastian;Grozhik, Anya V.;Olarerin-George, Anthony O.;Meydan, Cem;Mason, Christopher E.;Jaffrey, Samie R.
通讯作者：
Jaffrey, Samie R.
影响因子：
4.6
作者：
Lesbirel S;Viphakone N;Parker M;Parker J;Heath C;Sudbery I;Wilson SA
通讯作者：
Wilson SA
影响因子：
10.5
作者：
Ke S;Alemu EA;Mertens C;Gantman EC;Fak JJ;Mele A;Haripal B;Zucker-Scharff I;Moore MJ;Park CY;Vågbø CB;Kusśnierczyk A;Klungland A;Darnell JE Jr;Darnell RB
通讯作者：
Darnell RB