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The importance of mA topology in chicken embryo mRNA: a precise mapping of mA at the conserved chicken β-actin zipcode. | LitMetric
-methyladenosine (mA) in mRNA regulates almost every stage in the mRNA life cycle, and the development of methodologies for the high-throughput detection of methylated sites in mRNA using mA-specific methylated RNA immunoprecipitation with next-generation sequencing (MeRIPSeq) or mA individual-nucleotide-resolution cross-linking and immunoprecipitation (miCLIP) have revolutionized the mA research field. Both of these methods are based on immunoprecipitation of fragmented mRNA. However, it is well documented that antibodies often have nonspecific activities, thus verification of identified mA sites using an antibody-independent method would be highly desirable. We mapped and quantified the mA site in the chicken β-actin zipcode based on the data from chicken embryo MeRIPSeq results and our NA-pimodification etection and se-ecogniti (RedBaron) antibody-independent assay. We also demonstrated that methylation of this site in the β-actin zipcode enhances ZBP1 binding in vitro, while methylation of a nearby adenosine abolishes binding. This suggests that mA may play a role in regulating localized translation of β-actin mRNA, and the ability of mA to enhance or inhibit a reader protein's RNA binding highlights the importance of mA detection at nucleotide resolution.
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