Abstract
N6-methyladenosine (m(6)A) constitutes the most prevalent nucleotide modification within eukaryotic messenger RNA (mRNA). Variations in m(6)A levels are associated with numerous human diseases and health conditions, including various forms of cancer, diabetes, neurological disorders, male infertility, and obesity. Nevertheless, the molecular mechanisms underpinning the recognition of m(6)A by different 'reader' proteins remain incompletely elucidated. In this study, we used phage display to identify key sequence features that methyl readers recognize in m(6)A. This study shows that m(6)A modifications affect the mRNA interactome. A peptide motif recognizing m(6)A in DRACH sequences suggests a common recognition mechanism, though proteins may use different methods to detect m(6)A in less accessible areas. The sequence of the hnRNP A1 RRM domain that aligns with the newly discovered m(6)A-binding peptide, m1p1, is crucial for the binding of m(6)A-modified RNAs, indicating a strong link between the m1p1 sequence and m(6)A recognition, which is key for recognizing m(6)A-modified, unstructured RNAs. Gaining a comprehensive understanding of the evolutionary influence of m(6)A on its reader proteins may facilitate the identification of additional m(6)A readers. These signature peptides could enhance theranostic approaches across cancers, enabling more targeted therapies.