Mechanisms and Strategies for Determining m⁶A RNA Modification Sites by Natural and Engineered m⁶A Effector Proteins

Abstract

N6‐Methyladenosine (m6A) is the most common internal RNA modification in the consensus sequence of 5′‐RRACH‐3′. The methyl mark is added by writer proteins (METTL3/METTL14 metyltransferase complex) and removed by eraser proteins (m6A demethylases; FTO and ALKBH5). Recognition of this methyl mark by m6A reader proteins leads to changes in RNA metabolism. How the writer and eraser proteins determine their targets is not well‐understood, despite the importance of this information in understanding the regulatory mechanisms and physiological roles of m6A. However, approaches for targeted manipulation of the methylation state at specific sites are being developed. In this review, I summarize the recent findings on the mechanisms of target identification of m6A regulatory proteins, as well as recent approaches for targeted m6A modifications.

Crosstalk among N6-methyladenosine modification and RNAs in central nervous system injuries

“…It is an oxygenase dependent on Fe 2+ and 2-oxoglutarate, which can catalyze the demethylation of nucleotides ( Li L. et al, 2022 ). ALKBH5 is found after FTO, it is Fe 2+ and α- Ketoglutarate (α-Kg) dependent non-heme oxygenase ( Imanishi, 2022 ). Although both FTO and ALKBH5 are demethylases, they are differentially distribution in mammalian organs and have different substrate preferences.…”

The effect of METTL3 on MDM2 impairs cell cycle homeostasis in podocytes during diabetic kidney disease

“…m6A methyltransferases dynamically attach to the N6 position of the adenosine residues within the RRACH consensus sequence, forming m6A methylated RNA. This modi cation is particularly abundant around RNA termination codons, 3' UTR, and internal exons (15). In detail, heterodimer core complex of methyltransferase-like 3-methyltransferase-like 14 (METTL3-METTL14), and Wilms' tumor 1associating protein (WTAP) as a splicing factor, initiate modi cation of RNA as writers, while two main demethylases are fat mass and obesity-associated protein (FTO) and alkB homolog 5 (ALKBH5) as erasers.…”

High N6‐methyladenosine‐activated TCEAL8 mRNA is a novel pancreatic cancer marker

“…Many RNAs, including rRNA, U6 snRNA, mRNA, and ncRNA, undergo post‐transcriptional m6A functional modification in cells, and m6A modification is involved in the regulation of their functions and gene expression 2–4 . METTL3 has been identified as a representative m6A‐modifying enzyme and has been found to modify the central adenine of its binding sequence, RRACH (R: A or G, H: A, T, or C) 5 . The enzyme that performs the m6A modification is defined as a writer, the protein that binds to the m6A modification site is defined as a reader, and the enzyme that removes the m6A modification is defined as an eraser 6 .…”