Formation and determination of the oxidation products of 5-methylcytosine in RNA.
Formation and determination of the oxidation products of 5-methylcytosine in RNA.
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RNA 中 5-甲基胞嘧啶氧化产物的形成和测定
DOI：
10.1039/c6sc01589a
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发表时间：
2016-08-01
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：
Huang W;Lan MD;Qi CB;Zheng SJ;Wei SZ;Yuan BF;Feng YQ
化学标记与LC-MS相结合，能够灵敏地同时检测5-甲基胞嘧啶的氧化产物。用此方法可测定哺乳动物RNA中的5-甲基胞嘧啶、5-羟甲基胞嘧啶、5-甲酰胞嘧啶和5-羧基胞嘧啶。与DNA上可逆的表观遗传修饰类似，动态RNA修饰最近被认为以“RNA表观遗传学”的形式构成生物调控的另一个领域。5-甲基胞嘧啶（5-mC）长期以来一直被认为存在于所有三个生命王国的RNA中。然而，5-mC在RNA中的功能尚未完全了解，特别是对于RNA去甲基化机制。RNA中5-羟甲基胞嘧啶（5-hmC）的发现以及我们最近报道的RNA中5-甲酰胞嘧啶（5-foC）的发现表明RNA中5-mC可能经历与DNA中相同的胞嘧啶氧化去甲基化途径，通过10 - 11易位（泰特）蛋白产生中间体5-hmC、5-foC和5-羧基胞嘧啶（5-caC）。然而，到目前为止，尚未观察到RNA中的内源性5-caC。在本研究中，我们建立了一种方法，使用化学标记结合液相色谱-质谱分析的5-mC的氧化产物的灵敏和同时测定。结果表明，用2-溴-1-（4-二乙氨基苯基）-乙酮（BDEPE）标记后，RNA中5-mC、5-hmC、5-foC和5-caC的检测灵敏度提高了70-313倍。利用这种方法，我们发现哺乳动物RNA中存在5-caC。此外，我们发现5-mC存在于所有RNA种类，包括mRNA、28 S rRNA、18 S rRNA和小RNA（<200 nt）。而5-hmC、5-foC和5-caC主要存在于mRNA中，在其他类型的RNA中几乎检测不到。此外，我们还发现人结直肠癌（CRC）和肝细胞癌（HCC）组织RNA中5-hmC的含量较癌旁正常组织显著降低，提示RNA中5-hmC可能在肿瘤的发生、发展中起一定的调控作用。
Chemical labeling coupled with LC-MS enables the sensitive and simultaneous detection of the oxidative products of 5-methylcytosine. With this method, we can determine 5-methylcytosine, 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine in RNA of mammals. Similar to the reversible epigenetic modifications on DNA, dynamic RNA modifications were recently considered to constitute another realm for biological regulation in the form of “RNA epigenetics”. 5-Methylcytosine (5-mC) has long been known to be present in RNA from all three kingdoms of life. However, the functions of 5-mC in RNA have not been fully understood, especially for the RNA demethylation mechanism. The discovery of 5-hydroxymethylcytosine (5-hmC) in RNA together with our recently reported 5-formylcytosine (5-foC) in RNA indicated that 5-mC in RNA may undergo the same cytosine oxidation demethylation pathway with generating intermediates 5-hmC, 5-foC, and 5-carboxylcytosine (5-caC) by ten–eleven translocation (Tet) proteins as that in DNA. However, endogenous 5-caC in RNA has not been observed so far. In the current study, we established a method using chemical labeling coupled with liquid chromatography-mass spectrometry analysis for the sensitive and simultaneous determination of the oxidative products of 5-mC. Our results demonstrated that the detection sensitivities of 5-mC, 5-hmC, 5-foC and 5-caC in RNA increased by 70–313 folds upon 2-bromo-1-(4-diethylaminophenyl)-ethanone (BDEPE) labeling. Using this method, we discovered the existence of 5-caC in the RNA of mammals. In addition, we found the 5-mC occurs in all RNA species including mRNA, 28S rRNA, 18S rRNA and small RNA (<200 nt). However, 5-hmC, 5-foC and 5-caC mainly occur in mRNA, and barely detected in other types of RNA. Furthermore, we found that the content of 5-hmC in the RNA of human colorectal carcinoma (CRC) and hepatocellular carcinoma (HCC) tissues significantly decreased compared to tumor adjacent normal tissues, suggesting that 5-hmC in RNA may play certain functional roles in the regulation of cancer development and formation.
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影响因子：
64.5
作者：
Wu H;Zhang Y
通讯作者：
Zhang Y
DOI：
10.1126/science.1170116
发表时间：
2009-05-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Tahiliani M;Koh KP;Shen Y;Pastor WA;Bandukwala H;Brudno Y;Agarwal S;Iyer LM;Liu DR;Aravind L;Rao A
通讯作者：
Rao A
DOI：
10.1038/nrg2719
发表时间：
2010-03
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
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化学下拉揭示了哺乳动物转录组的动态假尿苷化
影响因子：
14.8
作者：
Li, Xiaoyu;Zhu, Ping;Yi, Chengqi
通讯作者：
Yi, Chengqi
哺乳动物 DNA 中 Tet 介导的 5-羧基胞嘧啶的形成及其 TDG 的切除
DOI：
10.1126/science.1210944
发表时间：
2011-09-02
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
He YF;Li BZ;Li Z;Liu P;Wang Y;Tang Q;Ding J;Jia Y;Chen Z;Li L;Sun Y;Li X;Dai Q;Song CX;Zhang K;He C;Xu GL
通讯作者：
Xu GL