The RNA m5C Methylase NSUN2 Modulates Corneal Epithelial Wound Healing.
The RNA m5C Methylase NSUN2 Modulates Corneal Epithelial Wound Healing.
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DOI：
10.1167/iovs.64.3.5
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发表时间：
2023-03-01
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
--
作者：
新兴的epitranscriptomics提供了各种RNA修饰的生理病理作用的见解。RNA甲基化酶N 0 P2/Sun结构域家族成员2（NSUN 2）催化mRNA的5-甲基胞嘧啶（m5 C）修饰。然而，NSUN 2在角膜上皮伤口愈合（CEWH）中的作用仍然未知。在这里，我们描述了NSUN 2在介导CEWH的功能机制。使用RT-qPCR、Western印迹、斑点印迹和ELISA来确定CEWH期间NSUN 2表达和总体RNA m5 C水平。进行NSUN 2沉默或过表达以探索其在体内和体外的CEWH中的参与。整合多组学以揭示NSUN 2的下游靶标。MeRIP-qPCR、RIP-qPCR和荧光素酶检测以及体内和体外功能检测阐明了CEWH中NSUN 2的分子机制。CEWH期间NSUN 2表达和RNA m5 C水平显著升高。NSUN 2敲低显著延迟体内CEWH并抑制体外人角膜上皮细胞（HCEC）的增殖和迁移，而NSUN 2过表达显著增强HCEC的增殖和迁移。从机制上讲，我们发现NSUN 2通过结合RNA m5 C阅读器Aly/REF输出因子来增加含有PHD和RING指结构域1（UHRF 1）的泛素样翻译。因此，UHRF 1敲低显著延迟体内CEWH，并抑制体外HCEC增殖和迁移。此外，UHRF 1过表达有效地挽救了NSUN 2沉默对HCEC增殖和迁移的抑制作用。NSUN 2介导的UHRF 1 mRNA的m5 C修饰调节CEWH。这一发现强调了这种新的epitranscriptomic机制在控制CEWH中的至关重要性。
The emerging epitranscriptomics offers insights into the physiopathological roles of various RNA modifications. The RNA methylase NOP2/Sun domain family member 2 (NSUN2) catalyzes 5-methylcytosine (m5C) modification of mRNAs. However, the role of NSUN2 in corneal epithelial wound healing (CEWH) remains unknown. Here we describe the functional mechanisms of NSUN2 in mediating CEWH. RT-qPCR, Western blot, dot blot, and ELISA were used to determine the NSUN2 expression and overall RNA m5C level during CEWH. NSUN2 silencing or overexpression was performed to explore its involvement in CEWH both in vivo and in vitro. Multi-omics was integrated to reveal the downstream target of NSUN2. MeRIP-qPCR, RIP-qPCR, and luciferase assay, as well as in vivo and in vitro functional assays, clarified the molecular mechanism of NSUN2 in CEWH. The NSUN2 expression and RNA m5C level increased significantly during CEWH. NSUN2 knockdown significantly delayed CEWH in vivo and inhibited human corneal epithelial cells (HCEC) proliferation and migration in vitro, whereas NSUN2 overexpression prominently enhanced HCEC proliferation and migration. Mechanistically, we found that NSUN2 increased ubiquitin-like containing PHD and RING finger domains 1 (UHRF1) translation through the binding of RNA m5C reader Aly/REF export factor. Accordingly, UHRF1 knockdown significantly delayed CEWH in vivo and inhibited HCEC proliferation and migration in vitro. Furthermore, UHRF1 overexpression effectively rescued the inhibitory effect of NSUN2 silencing on HCEC proliferation and migration. NSUN2-mediated m5C modification of UHRF1 mRNA modulates CEWH. This finding highlights the critical importance of this novel epitranscriptomic mechanism in control of CEWH.
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DOI：
10.1016/j.jtos.2021.03.005
发表时间：
2021-07
期刊：
The ocular surface
影响因子：
--
作者：
Koujah L;Allaham M;Patil CD;Ames JM;Suryawanshi RK;Yadavalli T;Agelidis A;Mun C;Surenkhuu B;Jain S;Shukla D
通讯作者：
Shukla D
影响因子：
3.4
作者：
Lavker RM;Kaplan N;Wang J;Peng H
通讯作者：
Peng H
影响因子：
11.4
作者：
Kan, Ryan L.;Chen, Jianjun;Sallam, Tamer
通讯作者：
Sallam, Tamer
影响因子：
4.5
作者：
Blanco S;Kurowski A;Nichols J;Watt FM;Benitah SA;Frye M
通讯作者：
Frye M
RNAm5Cfinder：基于随机森林预测 RNA 5-甲基胞嘧啶 (m5C) 位点的网络服务器
影响因子：
4.6
作者：
Li J;Huang Y;Yang X;Zhou Y;Zhou Y
通讯作者：
Zhou Y