乙酰化修饰降低Ku蛋白的DNA结合活性

Acetylation reduced the DNA binding activity of Ku protein

摘要:

[目的]研究乙酰化修饰对Ku蛋白活性的影响。[方法]利用耻垢分枝杆菌为表达菌株，转入Ku蛋白表达质粒，纯化具有乙酰化修饰的Ku蛋白和无乙酰化的Ku蛋白突变体，比较两类蛋白的生化活性；分析氧化压力和酸性环境下耻垢分枝杆菌细胞内Ku蛋白乙酰化水平的变化。[结果]Ku蛋白过量表达的耻垢分枝杆菌比转入空质粒的对照菌株生长缓慢；乙酰化Ku蛋白比未发生乙酰化Ku蛋白修复断裂DNA的活性降低、DNA结合活性降低；氧化压力和酸性压力环境下，耻垢分枝杆菌细胞内Ku蛋白数量降低，乙酰化Ku蛋白数量变化不大。[结论]乙酰化修饰能够调节Ku蛋白的DNA结合活性，从而调节非同源末端连接修复系统的活性；Ku蛋白乙酰化程度升高是耻垢分枝杆菌对不良生长环境的反应。

关键词:乙酰化;Ku;非同源末端连接修复途径;分枝杆菌

Abstract:

[Objective] To understand the role of acetylation modification on the activity of Ku protein.[Methods] Acetylated Ku protein and its non-acetylated mutant were expressed and purified using M smegmatis expression system, and then their biochemical activities were compared. The effect of oxidative stress and acidic environment on Ku protein acetylation level were analyzed in M. smegmatis. [Results] Ku protein over-expression M. smegmatis strain grew slower than the control strain transformed with the empty plasmid. Acetylated Ku protein had lower DNA repair activity and DNA binding activity than the non-acetylated Ku. Quantity of Ku protein in M. smegmatis cells under oxidative and acidic stress decreased, whereas there was subtle change of acetylated Ku protein. [Conclusion] Acetylation modification can regulate the DNA binding activity of Ku protein, thus regulate the activity of Non-homologous end joining system. The increase of acetylation level of Ku protein is the response of mycobacteria against the adverse growth environment.

Key words:acetylation;Ku;NHEJ;mycobacteria