| Avian leucosis used to cause huge lossed in our country,but now has been listed in the Medium and Long Term Planning of National Animal Disease Prevention and Control(2012-2020).Except for 11 subgroups(A to K),the high variability can lead to recombination and form new subgroup.Epidemiological investigation shows that subgroup J,subgroup A and subgroup B have been the common exogenous ALV,and resulted in serious economic losses.The study of the infection and tumorigenic mechanism of ALV is beneficial for the control and eradication of this disease.An ALV-B/ALV-J chimera CD08 isolate from hemangioma case that can cause cytopathic effection(CPE)in DF-1 cell was used in this study,and this virus was recombinant with ALV-B enevelope gene and ALV-J genomic frame.Based on the infectious molecular cloning of the ALV-B/ALV-J chimera virus,iTRAQ LC-MS/MS technology was used to detect the significantly changed proteins in two different time points(60hpi and 108hpi)with rescued virus,which was predicted to reveal the mechanism of ALV infection and CPE of ALV-B/ALV-J at molecular level.The rescued virus rCD08 has the similar CPE with its parent virus and is genetically stable during passage.The result of iTRAQ LC-MS/MS showed that 4444 proteins were detected in the infected samples.The differently expressed proteins were revealed during compared to COG database,among which 796 proteins were functional proteins;368 proteins involved in posttranslational modification,and protein turnover and chaperones;296 proteins for ranslation and ribosomal structure;236 proteins for replication,repair and recombination;and the rest of the proteins were related to transcription,signal transduction mechanisms,energy production and conversion,metabolism and intracellular trafficking.Analyzed by IPA,the differently expressed proteins in the cells at the earlier time point got involved in 60 pathways and 501 functions;and the changed proteins in the cells at the later time point got involved in 80 pathways and 501 functions.Meanwhile,we also found that the proteins like CAV1,CLTA and CLTB were related to viral invasion,and CAV1 might related to the mechanism of tumor;HMGB1 was down regulated in this study,MIF,FADD,GRB2,MMP2 and MAP2K1 were found in this study which related to the tumor.What's more,the up regulated of DJ-1 which was related to tumor was found in this study.In addition,in the cells which had the obvious CPE,the differently expressed proteins were related to EIF pathway which was activated and Paxillin pathway which was inhibited,and the proteins like FADD,MAPK14 and HMGB1 were related to CPE.The data and the result of this study can support the further study in exploring the pathogenesis ALV-B/ALV-J chimera virus and molecular mechanism of CPE. |
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