Mechanism Of M6A RNA Methylation Affecting α-synuclein Expression

Parkinson’s disease is a kind of neurodegenerative disease which is more common in middle-aged and elderly population.The characteristic marker in the brain of PD patients is the appearance of lewy bodies with alpha-synuclein(SNCA)as the main component.More and more evidence shows that epigenetic modification plays an important role in PD development processes,such as DNA methylation modification,histone acetylation modification and chromatin remodeling.N6-methyladenosine(m6A)RNA methylation modification is widely present in m RNA of various eukaryotes and other non-coding RNAs,and may be involved in the transcription,processing,translation,degradation and other processes.At present,the role of RNA methylation modification in the pathogenesis of PD is still unclear.Studies have shown that m6 A demethylase FTO can cause intracellular mitochondrial dysfunction and increased oxidative stress leading to neuronal cell apoptosis.This subject mainly studies the molecular mechanism of m6 A RNA methylation to regulate the expression of PD key protein(α-synuclein).In this study,we first constructed a PC12 cell line that stably knocked down m6 A demethylase FTO or ALKBH5,and treated cells with m6 A methylation inhibitor(cycloleucine).The results showed that m6 A RNA methylation was altered in the cell The modified content will significantly inhibit the transcription and translation levels of SNCA.Secondly,we studied the m6 A modification site on SNCA m RNA by online prediction and immunity;after knocking down the m6 A recognition proteins YTHDF1 and YTHDF2,we explored the effect of m6 A modification on α-synuclein expression.The results showed that there are two m6 A sites in the CDS region of SNCA m RNA.Knocking down the demethylase may directly affect the expression of SNCA gene,and increase the expression of α-synuclein after inhibiting the expression of YTHDF2,indicating that m6 A regulatory α-synuclein expression is achieved by YTHDF2 promoting its degradation.Finally,we examined the effects of m6 A on cell autophagy and its AKT / m TOR signaling pathway by detecting the difference in the expression of α-synuclein intracellular and extracellular.TThe experimental results show that after knocking down FTO and ALKBH5,the expression of p-AKT and p-m TOR protein increases while the expression of autophagy marker protein LC3Ⅱdecreases,and the expression of p62 increases,indicating that enhanced m6 A methylation can activate the AKT / m TOR pathway,thereby inhibiting cell Autophagy activity and ultimately regulates the transfer and extracellular release of α-synuclein.In summary,m6 A RNA methylation modification can inhibit the expression of SNCA m RNA by recognizing protein YTHDF2 regulating stability,and further affect the extracellular secretion of α-synuclein by affecting the AKT/m TOR signaling pathway to inhibit the occurrence of autophagy.This study reveals the molecular mechanism by which m6 A RNA methylation modification regulates the expression of key proteins(α-synuclein)in parkinson’s disease,and provides support and reference for RNA epigenetic modification in parkinson’s disease pathogenesis and its treatment.