| Rabies is a fatal neurological disease caused by rabies virus(RABV).Usually,rabies is prevented and controlled through vaccination of domestic dogs and cats,but it still causes nearly 5900 deaths worldwide each year,especially in developing countries where domestic vaccination rates are low.Almost all mammals are susceptible to rabies,and once clinical symptoms appear,it is almost 100%fatal.Rabies can be transmitted to humans from infected animals(mainly domestic dogs)through bites or scratches through saliva.Rabies can be prevented by vaccination before or after exposure,but there is no antivirus that can directly target the virus drug.Part 1:Ubiquitination is an important post-translational modification(PTM),which involves a variety of biological functions,and many studies have shown that it also plays a key role in viral infection.We performed ubiquitination studies on the brains of RABVinfected mice,aiming to provide new research materials for the pathogenesis of rabies virus by revealing the differences in the overall levels of ubiquitinated proteins in the brain tissue of infected mice.1 A quantitative ubiquitomic study was performed on RABV strain CVS-11-infected mouse brains to detect global changes in ubiquitination in mice brain tissues caused by rabies virus infection.Besides,we verified our experimental data and found that it met the quality standards;2 A total of 4,243 ubiquitinated sites were identified and these modification sites were distributed on 1,626 host proteins;these ubiquitination modification levels were screened using a threshold of 2-fold change,and a total of 644 and 70 ubiquitination Proteins undergo up-and down-regulation of modification levels,respectively were found;3 Combined with bioinformatics,the identified differentially ubiquitinated modified proteins were further analyzed.GO analysis showed that the differentially ubiquitinated proteins(DUPs)were significantly enriched in the myelin sheath of the cell and the binding activity;KEGG pathway analysis suggested that the identified proteins were mainly related to the biosynthesis of amino acids;4 It is observed the ubiquitination of all five RABV proteins in the data.We verified by co-immunoprecipitation and found that they are consistent with the results of ubiquitomic data.Part 2:During the transcription of the genome,only about 2%of the genes have the function of encoding proteins.The rest of the non-coding RNAs were initially thought to be the "noises" without biological function during genome transcription,including lncRNA,which is a by-product of RNA polymerase Ⅱ transcription.However,recent studies have shown that lncRNAs are involved in many important regulatory processes such as X chromosome silencing,genomic imprinting and chromatin modification,transcriptional activation,transcriptional interference,and nuclear transport.These regulatory effects of lncRNA have begun to cause widespread attention.In this section,we research the data from microarray and hope to find lncRNAs that regulate RABV infection.1 Verify and screen lncRNAs that are differentially expressed in the data,and obtain 7 lncRNAs that are up-regulated and consistent with the omics data;2 The lncRNA obtained above were objected to overexpression screening and found that lncRNA-ZNF582 can significantly inhibit the replication of RAB V;3 The inflammatory factors regulated by lncRNA-ZNF582 were detected and found that IFN-β,IL-1β and TNF-α were significantly inhibited;4 In combination with bioinformatic methods,a preliminary screening of lncRNAZNF582 target genes was performed.The four target genes CCNE1,PDGFD,ATF4,XIAP and PLCB4 were found to significantly down-regulated in host cells overexpressing lncRNAZNF852. |
