Identify The Recognition Protein In The Process Of FTO Modulating MZF1 Through M~6A Modification

RNA modifications are common in eukaryotes,includingRNA methylation,acetylation,uridine,etc.Among the more than 150RNA modifications that have been identified so far,the proportion of N6-methyladenosine(m~6A)maximum.RNA methylation mainly occurs in the motif of RRACH,which is enriched in long exons around and inside 3'UTR,stop codons,andRNA methylation also occurs in precursorRNAs(pre-RNAs)and long exons.Strands of non-codingRNAs(lncRNAs).In the process of regulatingRNA methylation,it is mainly regulated by three core components,namely:RNA methyltransferase,demethylase and recognition protein,which can add,delete and recognize the modification position of m~6A However,the RRm~6ACH motifs on different genes exert different binding effects to achieve different functions.This indicates that the function of m~6A modification depends not only on the specific RRACH sequence of m~6A,but also on the flanks of the m~6A site.The base sequence is related.The m~6A modification has been proved to be closely related toRNA expression,translation,splicing and stability,and is related to a variety of cancers,as well as to the prognosis and survival time of cancer patients.FTO is the first m~6A demethylase identified and is involved in a variety of biological functions.In the previous research basis,the candidate target gene MZF1 of FTO was screened,and the two m~6A modification sites and sequences on MZF1 were deter mined.However,the recognition protein that functions in this regulation process is still unclear.On this basis This study used Hela cells transiently transfected with YTHDF2 interferingRNA and overexpression vector as materials to verify the regulatory effect of YTHDF2 on MZF1 from the post-transcriptional level,and studied the effect of YTHDF2 on mRNA stability and ter mination of MZF1.The two potential m~6A sites near the codon were mutated,and the dual-luciferase reporter experiment was used to deter mine the m~6A site and sequence that played a regulatory role in the process of YTHDF2 regulating MZF1,as well as the interaction between the two sites,and explore the interference The effect of YTHDF2 on cell migration,proliferation and other phenotypes preli minarily reveals the effect of YTHDF2 on Hela cell biology by recognizing the m~6A modification site of MZF1.