Anti-sense expression of a metallopeptidase gene enhances nuclear entry of HBV-DNA.
Although several putative hepatitis B virus (HBV) receptors have been identified, none of them is capable of initiating HBV replication in a non-permissive human cell line. Using an Epstein-Barr virus-based extrachromosomal replication system, we have screened through a human liver cDNA library and successfully identified a clone capable of facilitating nuclear transport of HBV-DNA during the early phase of HBV infection. This clone contained a cDNA encoding a metallopeptidase-like protein in anti-sense orientation. Pretreatment of naïve HepG2 cells with 1,10-phenanthroline, an inhibitor for liver metallopeptidases, led to nuclear entry of HBV-DNA after HBV infection. However, cccDNA was still undetectable in the nuclei, indicating other cellular factors required to complete the replication cycle were still missing. Our present data suggest that in the initial stage of HBV infection, liver metallopeptidase constitutes a barrier for effective nuclear entry of HBV genomic DNA. Attenuation of metallopeptidase activity may facilitate HBV infection.
Yeh
Chau-Ting
CT
Liver Research Unit, Chang Gung Memorial Hospital and Chang Gung University, College of Medicine, Taipei, Taiwan, ROC. chauting@adm.cgmh.org.tw
Lai
Hsin-Yu
HY
Chu
Shiou-Ping
SP
Tseng
I-Chu
IC
D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D018076:DNA, Complementary / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics; D015723:Gene Library; D005821:Genetic Techniques; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D045726:Metalloproteases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008957:Models, Genetic; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D010618:Phenanthrolines / Q000494:pharmacology; D012313:RNA / Q000737:chemistry; D014162:Transfection
D018076:DNA, Complementary; D004279:DNA, Viral; D016376:Oligonucleotides, Antisense; D010618:Phenanthrolines; D012313:RNA; D004247:DNA; D045726:Metalloproteases; C025205:1,10-phenanthroline