Novel dual methylation of cytidines in the RNA of mammals.
Novel dual methylation of cytidines in the RNA of mammals.
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哺乳动物RNA中胞苷的新型双甲基化
DOI：
10.1039/d1sc01972d
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发表时间：
2021-05-07
期刊：
影响因子：
8.4
通讯作者：
Feng YQ
中科院分区：
文献类型：
--
作者：
Cheng MY;You XJ;Ding JH;Dai Y;Chen MY;Yuan BF;Feng YQ
RNA修饰在调节多种生理过程中起着关键作用。甲基化是发生在RNA中最普遍的修饰。在哺乳动物中，已报道了3种异构体胞苷甲基化修饰，包括3-甲基胞苷(M3C)、N4-甲基胞苷(M4C)和5-甲基胞苷(M5C)。除了胞苷碱基上的单一甲基化外，核糖的2‘羟基和胞苷碱基上的双重甲基化修饰也被报道过，包括N4，2’-O-二甲基胞苷(M4Cm)和5，2‘-O-二甲基胞苷(M5Cm)。在大肠杆菌的16S rRNA中发现了m4Cm，而在终末嗜热古菌和哺乳动物的tRNA中发现了m5Cm。然而，与m4Cm和m5Cm不同的是，推测的3，2‘-O-二甲基胞苷(M3Cm)的双重甲基化在生物体中从未被发现。因此，m3Cm在RNA中的存在仍然是一个悬而未决的问题。在本研究中，我们合成了m3Cm，并建立了LC-ESI-MS/MS方法来测定胞苷、m3Cm、m4Cm和m5Cm的二甲基化。在优化的分析条件下，可以清晰地区分m3Cm、m4Cm和m5Cm。利用这种方法，我们发现在哺乳动物的RNA中存在m3Cm。已鉴定的m3Cm是一种新的修饰，在包括古细菌、细菌和真核生物在内的三个结构域系统中尚未报道。我们证实，m3Cm主要存在于哺乳动物的小RNA(~lt；200nt)中。此外，我们还首次在哺乳动物细胞的18S rRNA中发现了m4Cm的存在。质谱监测的稳定同位素示踪表明，S-腺苷-L-蛋氨酸是核糖核酸中三种胞苷二甲基化的甲基供体。M3Cm的发现拓宽了生物体内RNA修饰的多样性。此外，m3Cm和m4Cm在哺乳动物中的发现为理解RNA修饰介导的RNA加工和基因表达调控开辟了新的方向。
RNA modifications play critical roles in regulating a variety of physiological processes. Methylation is the most prevalent modification occurring in RNA. Three isomeric cytidine methylation modifications have been reported in RNA, including 3-methylcytidine (m3C), N4-methylcytidine (m4C), and 5-methylcytidine (m5C), in mammals. Aside from the single methylation on the nucleobase of cytidines, dual methylation modifications occurring in both the 2′ hydroxyl of ribose and the nucleobase of cytidines also have been reported, including N4,2′-O-dimethylcytidine (m4Cm) and 5,2′-O-dimethylcytidine (m5Cm). m4Cm has been found in the 16S rRNA of E. coli, while m5Cm has been found in the tRNA of terminal thermophilic archaea and mammals. However, unlike m4Cm and m5Cm, the presumed dual methylation of 3,2′-O-dimethylcytidine (m3Cm) has never been discovered in living organisms. Thus, the presence of m3Cm in RNA remains an open question. In the current study, we synthesized m3Cm and established a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method to determine the dimethylation of cytidines, m3Cm, m4Cm and m5Cm. Under optimized analytical conditions, m3Cm, m4Cm and m5Cm can be clearly distinguished. Using the method, we discovered the existence of m3Cm in the RNA of mammals. The identified m3Cm is a novel modification that hasn't been reported in the three-domain system, including archaea, bacteria, and eukaryotes. We confirmed that m3Cm mainly existed in the small RNA (<200 nt) of mammals. In addition, we identified, for the first time, the presence of m4Cm in the 18S rRNA of mammalian cells. The stable isotope tracing monitored by mass spectrometry demonstrated that S-adenosyl-l-methionine was a methyl donor for all three dimethylations of cytidines in RNA. The discovery of m3Cm broadens the diversity of RNA modifications in living organisms. In addition, the discovery of m3Cm and m4Cm in mammals opens new directions in understanding RNA modification-mediated RNA processing and gene expression regulation.
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影响因子：
3.9
作者：
Guy MP;Shaw M;Weiner CL;Hobson L;Stark Z;Rose K;Kalscheuer VM;Gecz J;Phizicky EM
通讯作者：
Phizicky EM
DOI：
10.1002/cber.19480810406
发表时间：
1948-01-01
期刊：
CHEMISCHE BERICHTE-RECUEIL
影响因子：
--
作者：
BREDERECK, H;HAAS, H;MARTINI, A
通讯作者：
MARTINI, A
影响因子：
10.5
作者：
Ignatova, Valentina V.;Stolz, Paul;Schneider, Robert
通讯作者：
Schneider, Robert
DOI：
10.1016/j.jchromb.2004.10.051
发表时间：
2005-01-25
影响因子：
3
作者：
Liebich, HM;Müller-Hagedorn, S;Gérard, H
通讯作者：
Gérard, H
用于灵敏测定 RNA 中尿苷修饰的化学标记
影响因子：
8.4
作者：
Cheng QY;Xiong J;Ma CJ;Dai Y;Ding JH;Liu FL;Yuan BF;Feng YQ
通讯作者：
Feng YQ