CPSF30-L-mediated recognition of mRNA m6A modification controls alternative polyadenylation of nitrate signaling-related gene transcripts in Arabidopsis
CPSF30-L-mediated recognition of mRNA m6A modification controls alternative polyadenylation of nitrate signaling-related gene transcripts in Arabidopsis
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拟南芥中 CPSF30-L 介导的 mRNA m6A 修饰识别控制硝酸盐信号相关基因转录本的选择性多腺苷酸化
DOI：
10.1016/j.molp.2021.01.013
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发表时间：
2021-04-05
期刊：
影响因子：
27.5
通讯作者：
Li, Sisi
中科院分区：
文献类型：
--
作者：
Hou, Yifeng;Sun, Jing;Li, Sisi
N-6-甲基腺苷（m（6）A）是真核生物中普遍存在的一种mRNAs内部修饰，在mRNA代谢的各个方面都起着重要的作用。然而，很少有证据证明m（6）A在调节植物中的交替多聚腺苷酸化（阿帕）中的作用。阿帕是由一个大的蛋白质-RNA复合物与许多组件，包括裂解和多聚腺苷酸化特异性因子30（CPSF 30）的控制。在拟南芥中，CPSF 30具有两种同种型，并且较长的同种型（CPSF 30-L）含有植物所特有的YT 512-B同源（YTH）结构域。在本研究中，我们发现CPSF 30-L YTH结构域在体外与m（6）A结合。在cpsf 30 -2突变体中，包括几个重要的硝酸盐信号相关基因在内的许多基因的转录本中多聚腺苷酸化位点发生了与m（6）A峰相关的移位，表明这些携带m（6）A的基因转录本倾向于受阿帕的调控。野生型CPSF 30 L可以挽救cpsf 30 -2的阿帕和硝酸盐代谢缺陷，而m（6）A结合缺陷型CPSF 30-L突变体不能。综上所述，我们的研究结果表明，m（6）A修饰调节拟南芥中的阿帕，并揭示了m（6）A阅读器CPSF 30-L通过控制阿帕影响硝酸盐信号传导，为m（6）A修饰在硝酸盐代谢中的RNA 30端加工中的作用提供了新的线索。
N-6-methyladenosine (m(6)A), a ubiquitous internal modification of eukaryotic mRNAs, plays a vital role in almost every aspect of mRNA metabolism. However, there is little evidence documenting the role of m(6)A in regulating alternative polyadenylation (APA) in plants. APA is controlled by a large protein-RNA complex with many components, including CLEAVAGE AND POLYADENYLATION SPECIFICITY FACTOR30 (CPSF30). In Arabidopsis, CPSF30 has two isoforms and the longer isoform (CPSF30-L) contains a YT512-B Homology (YTH) domain, which is unique to plants. In this study, we showed that CPSF30-L YTH domain binds to m(6)A in vitro. In the cpsf30-2 mutant, the transcripts of many genes including several important nitrate signaling-related genes had shifts in polyadenylation sites that were correlated with m(6)A peaks, indicating that these gene transcripts carrying m(6)A tend to be regulated by APA. Wild-type CPSF30L could rescue the defects in APA and nitrate metabolism in cpsf30-2, but m(6)A-binding-defective mutants of CPSF30-L could not. Taken together, our results demonstrated that m(6)A modification regulates APA in Arabidopsis and revealed that the m(6)A reader CPSF30-L affects nitrate signaling by controlling APA, shedding new light on the roles of the m(6)A modification during RNA 30-end processing in nitrate metabolism.