鲍曼不动杆菌感染HeLa细胞与YY1互作的差异蛋白鉴定

Identification of differential proteins interacting with YY1 in HeLa cells infected by Acinetobacter baumannii

[目的]鉴定鲍曼不动杆菌感染HeLa细胞前后与YY1互作的差异蛋白。[方法]用脂质体转染法在HeLa细胞中过表达YY1;之后用Co-IP技术捕获与YY1相互作用的蛋白质;最后利用高效液相色谱质谱联用技术鉴定互作差异蛋白。用GO注释和KEGG分析预测蛋白质的功能。[结果]经Co-IP和蛋白银染发现鲍曼不动杆菌感染组和YY1捕获组与IgG组比较在45~55 kDa和70~100 kDa处有明显的差异条带;质谱结果显示YY1捕获组的特有蛋白有81个,GO注释和KEGG分析表明这些差异蛋白定位于细胞核及核膜,具有RNA结合功能,主要参与了RNA剪切的生物学过程;与IgG组和YY1捕获组比较在鲍曼不动杆菌感染组中获得了SFRS4、CPSF2、LUC7L2和U2AF65这些差异蛋白。GO注释和KEGG分析表明这些差异蛋白定位于细胞核散斑,具有RNA结合功能,主要参与了mRNA剪切和核输出的生物学过程。[结论]鲍曼不动杆菌感染的HeLa细胞中与YY1作用的蛋白均参与了RNA剪切过程,提示鲍曼不动杆菌很可能利用这些蛋白来调节YY1,为进一步研究鲍曼不动杆菌与YY1的相互作用机制奠定了基础。

[Objective] To identify the differential proteins interacting with YY1 before and after infection of HeLa cells by Acinetobacter baumannii.[Method] YY1 was overexpressed in HeLa cells by lipofection;Then, the protein interacting with YY1 was captured by Co-IP method;Finally, the interacting differential proteins were identified by high performance liquid chromatography-mass spectrometry.GO annotation and KEGG analysis were used to predict protein function.[Result] By Co-IP and silver staining, it was found that there were significant difference bands at 45-55 kDa and 70-100 kDa between Acinetobacter baumannii infection group and YY1 capture group and IgG group;The results of mass spectrometry showed that there were 81 unique proteins in YY1 capture group.GO annotation and KEGG analysis showed that these differential proteins were located in the nucleus and nuclear membrane, and had RNA binding function, mainly participated in the biological process of RNA splicing;Compared with IgG group and YY1 capture group, SFRS4,CPSF2,LUC7L2 and U2AF65 were obtained in Acinetobacter baumannii infection group.GO annotation and KEGG analysis showed that these differential proteins located in nuclear speckles, and had RNA binding function, mainly involved in the biological process of mRNA shearing and nuclear export.[Conclusion] In Acinetobacter baumannii infected HeLa cells, the proteins that interact with YY1 are involved in the RNA cleavage process.This suggesting that Acinetobacter baumannii may use these proteins to regulate YY1,which lays a foundation for further study on the interaction mechanism between Acinetobacter baumannii and YY1.