Direct RNA sequencing reveals m(6)A modifications on adenovirus RNA are necessary for efficient splicing.
Direct RNA sequencing reveals m(6)A modifications on adenovirus RNA are necessary for efficient splicing.
复制标题
DOI：
10.1038/s41467-020-19787-6
复制
发表时间：
2020-11-26
影响因子：
16.6
通讯作者：
Weitzman MD
中科院分区：
文献类型：
--
作者：
Price AM;Hayer KE;McIntyre ABR;Gokhale NS;Abebe JS;Della Fera AN;Mason CE;Horner SM;Wilson AC;Depledge DP;Weitzman MD
腺病毒是一种依赖于宿主RNA加工机制的核复制DNA病毒。细胞RNA的加工和代谢可以由胃L3调节，胃L3催化N6-甲基腺苷（m6 A）添加到mRNA。虽然先前已经检测到m6 A修饰的腺病毒RNA，但该标记在感染周期内的位置和功能尚不清楚。由于复杂的腺病毒转录组包括重叠的剪接单元，这将阻碍使用短读测序进行准确的m6 A定位，因此我们在此使用meRIP-seq和直接RNA长读测序的组合对腺病毒转录组内的m6 A进行分析，以产生核苷酸和转录物分辨的m6 A检测。尽管早期和晚期病毒转录物都含有m6 A，但m6 A书写者L3的缺失通过降低其剪接效率而特异性地影响病毒晚期转录物。这些数据展示了一种新的技术，在核苷酸分辨率的个别转录本中发现m6 A，并强调了m6 A在调节病毒病原体剪接中的作用。腺病毒转录物含有N6-甲基腺苷（m6 A）修饰。在这里，作者使用Illumina meRIP-Seq和纳米孔直接RNA测序分析了腺病毒mRNA上的m6 A修饰位点，并展示了m6 A在病毒晚期mRNA剪接中的作用。
Adenovirus is a nuclear replicating DNA virus reliant on host RNA processing machinery. Processing and metabolism of cellular RNAs can be regulated by METTL3, which catalyzes the addition of N6-methyladenosine (m6A) to mRNAs. While m6A-modified adenoviral RNAs have been previously detected, the location and function of this mark within the infectious cycle is unknown. Since the complex adenovirus transcriptome includes overlapping spliced units that would impede accurate m6A mapping using short-read sequencing, here we profile m6A within the adenovirus transcriptome using a combination of meRIP-seq and direct RNA long-read sequencing to yield both nucleotide and transcript-resolved m6A detection. Although both early and late viral transcripts contain m6A, depletion of m6A writer METTL3 specifically impacts viral late transcripts by reducing their splicing efficiency. These data showcase a new technique for m6A discovery within individual transcripts at nucleotide resolution, and highlight the role of m6A in regulating splicing of a viral pathogen. Adenovirus transcripts contain N6-methyladenosine (m6A) modification. Here the authors profile m6A modification sites on adenovirus mRNAs using Illumina meRIP-Seq and nanopore direct RNA sequencing, and showcase a role for m6A in splicing of viral late mRNAs.
登录
查看更多内容
影响因子：
48
作者：
Flusberg, Benjamin A.;Webster, Dale R.;Lee, Jessica H.;Travers, Kevin J.;Olivares, Eric C.;Clark, Tyson A.;Korlach, Jonas;Turner, Stephen W.
通讯作者：
Turner, Stephen W.
DOI：
10.1261/rna.065219.117
发表时间：
2018-03
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Darnell RB;Ke S;Darnell JE Jr
通讯作者：
Darnell JE Jr
影响因子：
16
作者：
Heinz S;Benner C;Spann N;Bertolino E;Lin YC;Laslo P;Cheng JX;Murre C;Singh H;Glass CK
通讯作者：
Glass CK
影响因子：
16.6
作者：
Chen, Kai;Lu, Zhike;Wang, Xiao;Fu, Ye;Luo, Guan-Zheng;Liu, Nian;Han, Dali;Dominissini, Dan;Dai, Qing;Pan, Tao;He, Chuan
通讯作者：
He, Chuan
影响因子：
4.5
作者：
Bresson SM;Hunter OV;Hunter AC;Conrad NK
通讯作者：
Conrad NK