The detection and functions of RNA modification m6A based on m6A writers and erasers

Zhang, Wei; Yang, Qian; Jia, Guifang

doi:10.1016/j.jbc.2021.100973

Cited by 56 publications

(32 citation statements)

References 166 publications

(381 reference statements)

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“…METTL3 was one of the key m 6 A methyltransferase which contained METTL3, WTAP, METTL14, and KIAA1429 ( Roundtree et al, 2017 ). M 6 A methyltransferase was involved in the post-transcriptional methylation of adenosine residues in the mRNA of eukaryotic cells to form N6-methyladenosine ( Zhang et al, 2021 ). In addition, studies have shown that METTL3 can directly act on the translation initiation machinery and promote m 6 A modification of mRNA, regardless of the activity of methyltransferase and m 6 A reader.…”

Section: Discussionmentioning

confidence: 99%

METTL3-Mediated m6A RNA Methylation of ZBTB4 Interferes With Trophoblast Invasion and Maybe Involved in RSA

Huang

Zhang

et al. 2022

Front. Cell Dev. Biol.

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N6-methyladenosine (m6A) was the most abundant modification of mRNA and lncRNA in mammalian cells and played an important role in many biological processes. However, whether m6A modification was associated with recurrent spontaneous abortion (RSA) and its roles were still unclear.Methods: Methylated RNA immunoprecipitation sequencing (MeRIP-Seq) was used to study the global m6A modification pattern in RSAs and controls. RNA sequencing (RNA-Seq) was used to study the level of global mRNA in two groups. Real-time quantitative PCR (RT-qPCR) was used to verify the level of mRNA of METTL3 and ZBTB4. MeRIP–qPCR was conducted to test the level of ZBTB4 m6A modification in two groups. In order to further explore whether ZBTB4 was the substrate of METTL3, the HTR-8/SVneo (HTR-8) cell line was selected for the knockdown and overexpression of METTL3. To study whether METTL3 regulated the ZBTB4 expression by recognizing ZBTB4 mRNA m6A motifs in coding sequences (CDS), dual-luciferase reporter assay was conducted. RNA stability assays using actinomycin D were conducted to study the RNA stability of the HTR-8 cell line with METTL3 overexpression and knockdown. To illustrate the role of METTL3 in the invasion of trophoblast, matrigel invasion assays and transwell migration assays were conducted using the HTR-8 cell line with METTL3 overexpression and knockdown.Results: A total of 65 genes were found with significant differences both in m6A modification and mRNA expression. We found m6A methyltransferase METTL3 was significantly down-regulated in the RSA group. Through gene function analysis, RT-qPCR, MeRIP–qPCR validation experiment, knockdown, and overexpression of METTL3 in the HTR-8 cell line, ZBTB4 was selected as one target of METTL3. Furthermore, we clarified that METTL3 regulated the expression of ZBTB4 by recognizing ZBTB4 mRNA m6A motifs in the CDS using the dual-luciferase reporter assay and METTL3 regulated the invasion of trophoblast by altering the stability and expression of ZBTB4 by RNA stability, matrigel invasion, and transwell migration assays.Conclusion: Our study revealed the mechanism by which METTL3 regulated the stability and expression of ZBTB4 and the trophoblast migration ability of RSA. A new perspective was provided for exploring the mechanism of embryonic development in RSA patients.

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Section: Discussionmentioning

confidence: 99%

METTL3-Mediated m6A RNA Methylation of ZBTB4 Interferes With Trophoblast Invasion and Maybe Involved in RSA

Huang

Zhang

et al. 2022

Front. Cell Dev. Biol.

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“…N 6 -methyladenosine (m 6 A) modification, the most frequent chemical modification in eukaryotic RNA, is involved in many biological processes including cancer progression [ 36 , 37 ]. Emerging studies highlighte that m6A modulates gene expression, thereby regulating tumor development aspects from cancer cell growth, metastasis, self-renewal of cancer stem cells, and apoptosis [ 38 , 39 ].…”

Section: Discussionmentioning

confidence: 99%

Stabilization of UCA1 by N6-methyladenosine RNA methylation modification promotes colorectal cancer progression

Jiang

et al. 2021

Cancer Cell Int

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Background UCA1 is frequently upregulated in a variety of cancers, including CRC, and it can play an oncogenic role by various mechanisms. However, how UCA1 is regulated in cancer is largely unknown. In this study, we aimed to determine whether RNA methylation at N6-methyladenosine (m6A) can impact UCA1 expression in colorectal cancer (CRC). Methods qRT-PCR was performed to detect the level of UCA1 and IGF2BP2 in CRC samples. CRISPR/Cas9 was employed to knockout (KO) UCA1, METTL3 and WTAP in DLD-1 and HCT-116 cells, while rescue experiments were carried out to re-express METTL3 and WTAP in KO cells. Immunoprecipitation using m6A antibody was performed to determine the m6A modification of UCA1. In vivo pulldown assays using S1m tagging combined with site-direct mutagenesis was carried out to confirm the recognition of m6A-modified UCA1 by IGF2BP2. Cell viability was measured by MTT and colony formation assays. The expression of UCA1 and IGF2BP2 in TCGA CRC database was obtained from GEPIA (http://gepia.cancer-pku.cn). Results Our results revealed that IGF2BP2 serves as a reader for m6A modified UCA1 and that adenosine at 1038 of UCA1 is critical to the recognition by IGF2BP2. Importantly, we showed that m6A writers, METTL3 and WTAP positively regulate UCA1 expression. Mechanically, IGF2BP2 increases the stability of m6A-modified UCA1. Clinically, IGF2BP2 is upregulated in CRC tissues compared with normal tissues. Conclusion These results suggest that m6A modification is an important factor contributing to upregulation of UCA1 in CRC tissues.

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“…In mammals, RNA m6A methylation is catalyzed by a polyprotein complex composed of METTL3, METTL14, Wilms’ tumor 1-associating protein (WTAP), the human homolog of Drosophila Virilizer (KIAA1429) [ 56 ], and several cofactors not yet identified [ 57 , 58 ]. Their functions are summarized in Supplementary Table S1 .…”

Section: M6a Methylation Is One Of the Defense Mechanisms Against Pla...mentioning

confidence: 99%

The Reversible Methylation of m6A Is Involved in Plant Virus Infection

Yue

Yao

Zhao

2022

Biology

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In recent years, m6A RNA methylation has attracted broad interest and is becoming a hot research topic. It has been demonstrated that there is a strong association between m6A and viral infection in the human system. The life cycles of plant RNA viruses are often coordinated with the mechanisms of their RNA modification. Here, we reviewed recent advances in m6A methylation in plant viruses. It appears that m6A methylation plays a dual role during viral infection in plants. On the one hand, m6A methylation acts as an antiviral immune response induced by virus infection, which inhibits viral replication or translation through the methylation of viral genome RNAs. On the other hand, plant viruses could disrupt the m6A methylation through interacting with the key proteins of the m6A pathway to avoid modification. Those plant viruses containing ALKB domain are discussed as well. Based on this mechanism, we propose that new strategies for plant virus control could be designed with competitive antagonists of m6A-associated proteins.

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The detection and functions of RNA modification m6A based on m6A writers and erasers

Cited by 56 publications

References 166 publications

METTL3-Mediated m6A RNA Methylation of ZBTB4 Interferes With Trophoblast Invasion and Maybe Involved in RSA

METTL3-Mediated m6A RNA Methylation of ZBTB4 Interferes With Trophoblast Invasion and Maybe Involved in RSA

Stabilization of UCA1 by N6-methyladenosine RNA methylation modification promotes colorectal cancer progression

The Reversible Methylation of m6A Is Involved in Plant Virus Infection

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