Binding and inhibition of adeno-associated virus Rep78 protein with hepatitis B virus C promoter / 中华肝脏病杂志

ABSTRACT

<p><b>OBJECTIVES</b>Adeno-associated virus (AAV) Rep78 is known for its inhibitory effects on replication of several viruses and oncogenes transformations. The study was to investigate the effect of Rep78 on hepatitis B virus C (HBV-C) gene and the mechanism of it.</p><p><b>METHODS</b>HBV-C promoter and HBV-C gene with its promoter were amplified by PCR and labeled with 32P-ATP. Electrophoretic mobility shift assay (EMSA) and in vitro transcription were utilized to detect the binding of MBP-Rep78 with HBV-C promoter and the transcription of HBV-C gene.</p><p><b>RESULTS</b>EMSA showed that by increasing the amount of Rep78 protein from 0.1 microg to 1.0 microg, the binding bands got stronger in a dose-dependent manner. In addition, Rep78 antibody was used to certify the specificity of this binding. The compound of Rep78, Rep78 antibody and HBV-C promoter were seen as super shift bands in EMSA. Meanwhile, HBV-C gene transcription was significantly inhibited by in vitro transcription which meant that Rep78 could not only bind with HBV-C promoter, but also could inhibit the transcription of HBV-C gene.</p><p><b>CONCLUSION</b>AAV Rep78 could inhibit the transcription of HBV-C gene through its binding with HBV-C promoter.</p>