Exploring The Molecular Mechanism Of Clr1 Protein In DNA Damage Response In Fission Yeast

DNA damage response is a series of reactions that the body makes in response to damage.The effective coordination of DNA damage response plays an important role in the normal development and survival of the body.Studying the molecular mechanism of DNA damage response will deepen the understanding of the occurrence and development mechanism of many human diseases,and provide new ideas and methods for the prevention and treatment of diseases.In this study,the mechanism of DNA damage response in eukaryotes was studied by using fission yeast as model organism.ATR/Rad3 protein is an crucial upstream protein kinase in DNA damage response,which is involved in multiple regulation of DNA damage response.In the context of Rad3 gene deletion,the mutant strains that have complement on drug-sensitive phenotypes of DNA damage were screened out,and then the relevant mutant genes were determined by deep sequencing of the mutant strains.One of the new genes that may be involved in DNA damage response was Clr1.The Clr1 protein is a transcriptional negative regulatory protein and has been reported to be involved in the assembly of heterochromatin.Through genetic experiments,this study preliminarily verified the correlation between Clr1 protein and DNA damage response,and Clr1 deletion can complement the sensitivity of Rad3 deletion to DNA damage drugs.Through biochemical experiments,it was proved that Clr1 protein was degraded by the ubiquitination pathway by the regulation of Rad3 protein in the DNA damage response.However,as a downstream protein of Rad3,Clr1 is not directly regulated by Rad3 protein phosphorylation in damage response.Through mass spectrometry identification,it was found that multiple sites of Clr1 protein were phosphorylated after DNA damage drug treatment.The specific functions of these sites and the specific molecular mechanism of Rad3 protein regulating Clr1 protein still need to be proved by further experiments.This study also found that Clr1 protein increased in S phase,suggesting that it may be involved in the replication.If fission yeast is used as a model organism,it will face the problem that the cell wall is difficult to remove during the research,and A.lutues is an actinomycete that can release lyticase to lyse the cell wall of fission yeast.By using high-throughput sequencing and bioinformatics,this paper first analyzed the whole genome sequence of Arthrobacter luteus,providing a basis for subsequent research on various aspects.