[Binding and inhibition of adeno-associated virus Rep78 protein with hepatitis B virus C promoter].
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
AAV Rep78 could inhibit the transcription of HBV-C gene through its binding with HBV-C promoter.
Yan
Zhong-yu
ZY
Liver Disease Research Center, Beijing Friendship Hospital, Capital University of Medical Sciences, Beijing 100050, China.
Cong
Min
M
Wang
Ping
P
Tang
Shu-zhen
SZ
Wang
Bao-en
BE
Jia
Ji-dong
JD
Liu
Yong
Y
Yong
Hong
H
D004268:DNA-Binding Proteins / Q000235:genetics*; D000229:Dependovirus / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics*
D004268:DNA-Binding Proteins; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2