tailieunhanh - Báo cáo khoa học: Fidelity of hepatitis B virus polymerase

Although efficient vaccines are available, chronic hepatitis B (HBV) infection poses a major health problem worldwide, and prolonged treatment of chronicallyinfected HBV patients with nucleoside analogs often results in , it is critical toevaluate the contribution of the HBV polymerase to mutations. FLAG-taggedwild-type (FPolE) andmutant (FPolE/D551A)HBV polymerases have been expressed in insect cells and purified. The purified FPolE showed DNA polymerase activity, but FPolE/D551A did not, implying that the activity was derived from FPolE | Eur. J. Biochem. 270 2929-2936 2003 FEBS 2003 doi Fidelity of hepatitis B virus polymerase Sung Gyoo Park1 Younhee Kim2 Esther Park1 Hyun Mi Ryu1 and Guhung Jung1 1 School of Biological Science Seoul National University Seoul department of Oriental Medicine Semyung University Checheon Chungbuk Korea Although efficient vaccines are available chronic hepatitis B HBV infection poses a major health problem worldwide and prolonged treatment of chronically infeeted HVV patients with nucleoside analogs often results in drugresistant HBV variants. Therefore it is critical to evaluate the contribution of the HBV polymerase to mutations. FLAG-tagged wild-type FPolE and mutant FPolE D551A HBV polymerases have been expressed in insect cells and purified. The purified FPolE showed DNA polymerase activity but FPolE D551A did not implying that the activity was derived from FPolE. No 3 fi 5 exonuclease activity was detected in FPolE. The fidelity of FPolE was invettigated and compared with that of HIV-1 RT which is highly error-prone. The fidelityof HBV polymerase seems to be achieved by increasing the Km for the dNTP being misinserted. The nucleotide misinsertion efficiency fl FPolE and HV-1 RT ranged from X 10 4 C T to X 10 3 G T and from X 10 4 C T to X 10 3 G T respectively and the overall misinsertion efficiency of HIV-1 RT was just higher than that of FPolE implying that HBV polymerase is fairly error-prone. Though HBV genetic mutation rate in replication is thought to be between those in RNA and DNA viruses our data shows that the rate of mutation byHBV polymerase is higher than the rate of genetic mutation in vivo. This may he a essutt from noire overlapping HBV genes in the HBV genome than that of other retroviruses. Keywords HBV polymerase HBV fidelity misinsertion mispair exonuclease. The hepatitis B virus HBV is a member of the hepadnavi-ridae a family of enveloped hepatotaipic DNA vhuises. The virus can .

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