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N6-methyladenosine (m6A) is a highly prevalent RNA modification in mRNA and non-coding RNAs that affects RNA splicing, translation, and stability, as well as the epigenetic effects of certain non-coding RNAs1.
Recent advances in sequencing technology have resulted in sensitive techniques that can be used to map m6A to the transcriptome of various model systems and cell types. Knowing the transcriptome-wide location of m6A has given a clearer indication of this marks many functions. We now have a much greater understanding of the biological and regulatory roles being played by this epigenetic mark, although it is also clear that there is still much left to uncover. The same advances in sequencing methods and technology have also allowed us to determine the readers (m6A-binding proteins), writers (methyltransferases), and erasers (demethylases) responsible for m6A's functions, methylation, and demethylation.
Here, we cover the essential m6A writers and erasers as well as some of the crucial readers of m6A and the epigenetic functions they serve.
m6A is dynamically regulated within the nucleus by a methyltransferase writer complex, which deposits m6A on mRNA, and m6A demethylases, which will remove the mark.
The idea that m6A can be added to mRNA and subsequently removed will be an important consideration for future m6A research. Dynamic changes in m6A levels across the transcriptome may point to new functions for this epigenetic mark.
Figure 1. m6A writers and erasers. Click on the image to download the interactive pathway.
Achieve highly reproducible results in your epigenetic research with our recombinant monoclonal antibodies for key m6A writers and erasers. All of these antibodies are knockout validated to ensure their specificity.
|Target name||Function||Location||Recommended abID||Application||Species|
|METTL3||Install m6A on RNA (catalytic subunit)||Nucleus||ab195352||Flow Cyt, WB, IHC-P, ICC/IF, IP||Mouse, Rat, Human|
|WTAP||Localize METTL3 and METTL4 to nuclear speckles||Nucleus||ab195380||ICC/IF, IP, Flow Cyt, IHC-P, WB||Human|
|ALKBH5||Demethylate m6A||Nucleus||ab195377||WB, IHC-P||Mouse, Rat, Human|
|FTO||Demethylate m6Am||Nucleus||ab126605||WB, IHC-P, ICC/IF||Human|
m6A exerts its effects on RNA by recruiting m6A-binding proteins called readers. Knowledge of m6A readers and their functions is rapidly increasing. More and more studies are revealing new m6A readers with specific roles for m6A in different cell types and model systems.
m6A-binding proteins will commonly contain a YTH (YT521-B homology) domain. RNA pulldown has shown that proteins containing a YTH domain are commonly m6A binders11. These different YTH domain proteins have since been shown to have a broad spectrum of roles associated with their ability to bind m6A. YTHDF1, working together with YTHDF3, is known to promote the translation efficiency of its target RNAs7,12. YTHDF2 is thought to play a role in mRNA stability13. YTHDC1 is implicated in m6A-mediated alternative splicing14. YTHDC2 has been shown to enhance the translation efficiency of its targets while decreasing target abundance15.
Not all m6A readers are YTH containing; they also include eukaryotic initiation factor 3 (eIF3), heterogeneous nuclear ribonucleoproteins (HNRNPs), and insulin-like growth factor 2 mRNA-binding proteins (IGF2BPs). eIF3 plays a role in translation preinitiation and has also been confirmed to bind preferentially to m6A within the 5'UTR of mRNA leading to enhanced translation16. Two HNRNP proteins, HNRNPC and HNRNPG, regulate the processing of m6A-containing RNA transcripts, harboring m6A as a structural switch to make those transcripts more accessible for binding7. IGF2BPs have been reported to promote the stability and storage of their target mRNAs in an m6A-dependent manner17.
Apart from being involved in RNA metabolism, m6A readers also participate in different biological processes, including tumorigenesis, hematopoiesis, virus replication, immune response, and adipogenesis18.
Figure 2. m6A readers and their functions. Click on the image to download the interactive pathway.
To help your study of m6A functions, we have compiled a list of our best-selling recombinant monoclonal antibodies for m6A readers.
|Target name||Function1,7||Location||Recommended abIDs||Tested applications||Species|
|YTHDF1||Regulation of mRNA stability, translation||Cytoplasm||ab252346||WB, IHC-P, Flow Cyt||Mouse, Rat|
|YTHDF2||Regulation of mRNA stability||Cytoplasm||ab220163||WB, IP||Mouse, Rat, Human|
|ab246514||Flow Cyt, WB, IHC-P, IP, ICC||Mouse, Rat, Human|
|YTHDF3||Regulation of mRNA stability, translation||Cytoplasm||ab220161||Flow Cyt, IP, WB, IHC-Fr, IHC-P||Mouse, Rat, Human|
|YTHDC1||Alternative splicing, lncRNA-mediated gene silencing||Nucleus||ab220159||WB, IHC-P||Mouse, Rat, Human|
|YTHDC2||Translation||Cytoplasm||ab220160||IHC-P, Flow Cyt, IP, WB||Mouse, Rat, Human|
|HNRNPC||Regulation of splicing, structure switching||Nucleus||ab133607||WB, IHC-P, ICC/IF||Mouse, Rat, Human|
|HNRNPG||Nucleus||ab190352||WB, IHC-P, ICC/IF||Mouse, Rat, Human|
|IGF2BP1||Regulation of mRNA stability and storage||Cytoplasm||ab184305||WB, IHC-P, IP||Human|
|IGF2BP2||Cytoplasm||ab124930||WB, IHC-P, ICC/IF||Mouse, Rat, Human|
|IGF2BP3||Cytoplasm||ab177477||WB, ICC/IF, IP, Flow Cyt||Mouse, Rat, Human|
Various methods have been developed to map m6A throughout the transcriptome effectively. Initial studies have used immunoprecipitation methods with highly specific m6A antibodies followed by next-generation sequencing. These methods are known as MeRIP-seq. MeRIP has since advanced, and it is now possible to carry out single-nucleotide resolution m6A mapping. This can be achieved using very small fragments of RNA (100–200 nts) in combination with immunoprecipitation. This method is known as miCLIP (m6A individual-nucleotide-resolution-crosslinking and immunoprecipitation), and it was first described in Linder et al. 2015 using the Abcam m6A antibody