Effects Of Human Bocavirus Noncoding RNA On NF-?B Signaling Pathway

Human Bocavirus(HBoV)was first detected in human nasopharyngeal secretions by Swedish scientists in 2005.Human Bocavirus(HBoV)was classified into parvovirus family,Bocavirus genus.Boka virus is the fourth most common respiratory virus after respiratory syncytial virus,adenovirus and rhinovirus.A large amount of research data shows that as much as half of the DNA in higher organisms can be transcribed into RNA,and the majority of these RNAs are non-coding rnas.Non-coding RNA can participate in the post-transcriptional regulatory process of eukaryotic gene expression.Can also participate in the regulation of a variety of life activities.At the same time,pathogenic microorganisms have also evolved various molecular mechanisms to antagonize host innate immunity,including the use of self-encoding ncRNA.Nuclear factor kappa-B(NF-?B)is a protein complex.NF-?B is found in almost all cells and is involved in responses to external stimuli such as cytokines,radiation,heavy metals,and viruses.NF-?B plays a key role in inflammatory and immune responses.Non-coding RNA in the body can be involved in a variety of regulation.We speculate that the non-coding RNA of the virus may also have an impact on the invasion or replication of the virus or on the body's natural immunity.This study focused on the effect of human boka virus non-coding RNA on NF-?B signaling pathway.1.The non-coding RNA of bocavirus can inhibit the NF-?B signaling pathwayWe used the dual luciferase reporter system combined with fluorescence quantitative PCR to verify the effect of boka virus non-coding RNA on NF-?B signaling pathway.It was found that the activation of NF-?B was inhibited by HBoV1 ncRNA transfection.Subsequently,the dual-luciferase reporter system was used to detect the effect of HBoV1 ncRNA on NF-?B related signal molecules IKK,IKK and TRAF2,and it was found that HBoV1 ncRNA did not affect NF-?B upstream signal molecules.Immunofluorescence technique was used to demonstrate that HBoV1 ncRNA does not affect the nuclear translocation of p65 protein.2.Proteins interacting with HBoV1 ncRNA were identified and analyzedBased on the specific binding pathway of biotin and streptavidin,the RNA pull down and protein spectrum detection techniques were continued to screen proteins binding to HBoV1 ncRNA.In this study,a variety of specific and non-specific proteins interacting with HBoV1 ncRNA were co-precipitated,among which 406 proteins specifically bound to HBoV1 ncRNA.By comparing the differences in mass spectrometric identification results and protein-related functions between the experimental group and the control group,a total of three related transcription factor proteins were screened,namely TAF15,TFCP2 and RTF1.It was analyzed that HBoV1 ncRNA may affect the activation of NF-?B signaling pathway by binding to transcription factors.