肿瘤U1 snRNA蛋白互作图谱及特征分析

doi:10.3969/j.issn.1009-976X.2023.02.012

摘要/Abstract

摘要： 目的利用CHIRP-MS技术系统地鉴定肿瘤细胞中U1 snRNA的互作蛋白,并建立U1 snRNA高通量互作蛋白图谱,研究U1 snRNA在肿瘤细胞中新的生物学功能和分子调控机制。方法通过带生物素标记的U1 snRNA探针捕获与U1 snRNA结合的互作蛋白,并分别取部分样品纯化蛋白和RNA,通过qPCR与跑胶银染确认探针富集效果,然后利用高效液相色谱-质谱联用技术鉴定获取的蛋白,最后利用生物信息学的方法进行GO、KEGG、蛋白复合体富集分析和PPI分析。结果我们共鉴定到了135个高可靠的U1 snRNA结合蛋白,其中包括60个已知的和75个新的U1 snRNA结合蛋白。通路富集等生物信息学分析显示U1 snRNA协同调控RNA剪接、胞内运输、定位、端粒和表观遗传等多个生物学过程。蛋白互作网络分析揭示ESWR1等肿瘤相关蛋白可能介导了U1 snRNA的肿瘤调控功能。结论本研究在国际上首次建立了U1 snRNA互作蛋白图谱,揭示了U1 snRNA在肿瘤细胞中的多种生物学功能,为进一步研究U1 snRNA参与肿瘤发生发展以及肿瘤预防和治疗提供了理论基础。

关键词: 肿瘤, U1 snRNA, CHIRP-MS

Abstract: Objective The interaction proteins of U1 snRNA in tumor cells were systematically identified by CHIRP-MS technology. We established a high-throughput interaction protein map of U1 snRNA to study the new biological function and molecular regulation mechanism of U1 snRNA in tumor cells.Methods The biotin-labeled U1 snRNA probe captured the interaction protein binding with U1 snRNA, and some samples were taken to purify the protein and RNA. First, the enrichment effect of the probe was confirmed by qPCR and gel silver staining. Then the obtained protein was identified by high-performance liquid chromatography-mass spectrometry. Finally, the GO, KEGG, protein complex enrichment analysisand PPI analysis were performed by bioinformatics. Results We have identified 135 highly reliable U1 snRNA binding proteins, including 60 known and 75 new U1 snRNA binding proteins. Bioinformatics analysis, such as pathway enrichment, showed that U1 snRNA coordinated and regulated multiple biological processes such as RNA splicing, intracellular transport, localization, telomere, and epigenetics. In addition, the protein-protein interaction network analysis revealed that tumor-related proteins such as ESWR1 may mediate the tumor regulatory function of U1 snRNA. Conclusion In this study, the U1 snRNA interaction protein map was established for the first time in the world, revealing the multiple biological functions of U1 snRNA in tumor cellsand providing a theoretical basis for further study of U1 snRNA's involvement in tumor development, prevention, and treatment.

Key words: tumor, U1 snRNA, CHIRP-MS

中图分类号:

R73-37

田彬, 杨兵, 马星宇, 杨芝芝, 廖建友. 肿瘤U1 snRNA蛋白互作图谱及特征分析[J]. 岭南现代临床外科, 2023, 23(02): 180-189.

TIAN Bin, YANG Bing, MA Xing-Yu, YANG Zhi-Zhi, LIAO Jian-You. Analysis of protein interaction patterns and characteristics of U1 snRNA in cancer[J]. Lingnan Modern Clinics In Surgery, 2023, 23(02): 180-189.

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