Research For Proteome And Acetylome Analysis Of DNA Repair Proteins In Early DNA Damage Based On Mass Spectrometry

Background and research purpose:The DNA repair pathway of eukaryotic cells is responsible for repairing the DNA damage induced by internal and external stressors,maintaining the stability of the genome,and reducing the risk of cell cancer.Acetylation is a kind of universally existed post-translational modification of protein in cells,which is dynamicallly regulated by acetylases(HATs)and deacetylases(HDACs)and can regulate the function of repair proteins through various mechanisms.Abnormal acetylation modification can lead to functional dysregulation of DNA repair protein and promote the occurrence and progression of cancer.Targeted regulation of the acetylation modification on repair proteins to achieve anti-tumor effects is considered to be a new direction of anti-tumor therapy with great potential.The early stage of DNA damage is an important node for initiating damage repair.To the date,proteins regulated by acetylation in the early DNA damage events are not fully understood.We intend to provide and analyze the changes in the expression level and acetylation of early repair proteins in the early response of DNA damage.Method:In this study,with the combination of anti-lysine acetylation antibody enrichment technology combined with high-resolution 4D-label ree quantitative proteomics technology,we completed the identification and quantification of the whole proteome and acetylome in 293T cells treated with epirubicin for 1 hour.Result:A total of 5526 quantifiable proteins and 6789 acetylation sites on 2400 proteins were identified,including 190 acetvlation sites on 50 repair proteins.Most repair proteins were not significantly upregulated or down-regulated in the expression level.Among the 50 repair proteins,42 lysine sites on 21 repair proteins and 24 lysine sites on 16 repair proteins were acetylated and deacetylated respectively after epirubicin treatment.7 repair proteins simultaneously contained both acetylated and deacetylated lysine sites.GTF2H2C,RAD51C and RAD 17 have been found to have acetylation modification for the first time.The regualatory potential of acetylation on the function of 17 repair proteins was predicted according to the GO function annotation.For further analysis,1 1 lysine sites with acetylated or deacetylated changes were located in the functional domain of 7 proteins under the conditions of Epirubicin treatment,which are high possibly to regulate the function of the domains,thereby regulating the function of the protein.It might help us to further explore the effects of acetylation changes on these proteins.Conclusion:In conclusion:according to the result of proteome and acetylome that the acetylation changes on repair proteins was earlier than the changes on the expression of proteins in cells at early phase of DNA damage,indicating that the acetylation modification was indeed possibly an important mechanism in the response to the early stress of DNA damage.