DNA甲基化及突变对KRAS启动子区G-四链体结构的影响
首发时间:2022-05-11
摘要:目的:研究K-RAS基因启动子区富G序列胞嘧啶甲基化或突变对其G-四链体结构的影响。方法:首先利用胞嘧啶突变为胸腺嘧啶确定关键胞嘧啶,然后将对应位点的胞嘧啶甲基化,再通过圆二色谱和离子淌度质谱对比甲基化前后G-四链体结构的差异。结果:圆二色谱结果显示,甲基化后该序列在260 nm处的吸收增加,而在290 nm处的吸收均降低,说明G-四链体结构有序性增加;离子淌度质谱分析则发现,甲基化序列的漂移时间明显小于野生型及突变序列,说明甲基化后G-四链体结构更加收敛有序。结论:胞嘧啶甲基化有利于提升K-RAS Mid GG-四链体正向平行结构的稳定性和有序性。
关键词: K-RAS 离子淌度质谱 圆二色谱 5-甲基胞嘧啶 G-四链体
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Effects of DNA methylation and mutations on the structure of the G-quadruplex in KRAS promoter region
Abstract:Purpose: To investigate the effects of 5-methylcytosine on the structure of the G-quadruplex in the promoter region of the K-RAS gene. Methods: The key cytosine was firstly dertermined by the mutation of cytosine to thymine, and then the cytosine at the corresponding site was methylated.Therefore the differences before and after G-quadruplex methylation were compared by circular dichroism and ion mobility mass spectrometry. Results: The results of circular dichroism showed that the absorption of the methylated sequence increased at 260 nm and decreased at 290 nm, indicating that the structural order of G-quadruplex increased; Ion mobility mass spectrometry found that the drift time of methylated sequence was significantly less than that of wild-type and mutant sequence, indicating that the G-quadruplex structure was more convergent and orderly after methylation. Conclusion: cytosine methylation is beneficial to improve the stability and order of the parallel G-quadruplexstructure from K-RASMid G.
Keywords: K-RAS Ion mobility mass spectrometry Circular dichroism 5-Methylcytosine G-quadruplex
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DNA甲基化及突变对KRAS启动子区G-四链体结构的影响
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