The Interaction Of Hepatitis B X Protein HBx And TOMM70A And The Effects On Its Function In Hepatocytes

Hepatitis B virus(HBV)is still an important pathogen affecting the health of our people.Chronic HBV infection is closely related to the development and progression of severe liver disease such as liver cirrhosis and liver cancer.The risk of developing hepatocellular carcinoma(HCC)is much higher than that of patients without HBV infection,and it is regrettable that The specific mechanism of its pathogenesis is not yet fully clear.The HBV genome is a partially double-stranded circular DNA with a total length of about 3.2 kb,including four open reading frames(ORF): P,S,C,X;where the X protein(HBx)X protein coding.HBx molecular weight of 16.5kD,containing 154 amino acids,multifunctional protein,in the process of HBV infection plays an important role in HBV is one of the important pathogenic factors in the persistent infection of the virus,the host cell apoptosis,The occurrence and development of liver cancer have played a very important role in the process.Current studies indicate that the HBx protein interacting with a number of host factors,including transcriptional factors and molecules involved in intracellular signaling and apoptosis.Mitochondrial Epithelin TOMM70A(also known as TOM70)is one of the important constituent subunits of mitochondrial outer membrane TOM complex and is a gated channel for mitochondrial precursor protein transport.The TOM complex mainly recognizes and transports the mitochondrial precursor protein,which processes the mature mitochondrial proteins by processing the mitochondrial related proteins and then transfers them to the outer membrane,intima,membrane gap and matrix of the mitochondria.Because TOMM70 A is involved in the composition of the TOM complex is the majority of mitochondrial protein transport of the key channel,so the abnormal function may lead to obstruction of mitochondrial protein may lead to the occurrence of various diseases.In recent years,mitochondrial epithelial translocation enzyme complex in the body of innate immunity,cardiovascular disease and neurodegenerative diseases play an important role.In this study,we used yeast two-hybrid system to screen the hepatocyte protein interacting with HBx protein to obtain the mitomolytic membrane transporter TOMM70 A.On the basis of this,it was further confirmed that HBx protein could interact with TOMM70 A in hepatoma cell line HepG2,and to explore the biological significance of HBx and TOMM70 A interaction,and to further understand the pathogenesis of HBV.In the first part of this study,HBx expression vector pHBx-Flag and transfection of HepG2 cells on the basis of CytoTrap yeast two-hybrid preliminary screening results further confirmed that HBx and TOMM70 A were confirmed by co-immunoprecipitation There is interaction in HepG2 cells.The second part of the study was designed to investigate whether HBx interacts with TOMM70 A for TOMM70 A and its related functions.As a result of the literature reported that TOMM70 A and Hsp90 / TBK1 / IRF3 interaction and then induced type I interferon.The effect of HBx expression on the interaction of TOMM70 A with Hsp90 / TBK1 / IRF3 was compared by immunoprecipitation technique.The results showed that HBx expression could affect the interaction between TOMM70 A and Hsp90 / TBKI/1IRF3.This suggests that HBx may affect the induction of type I interferon by competing with TOMM70 A to influence the interaction of Hsp90 / TBK1 / IRF3 with TOMM70 A.