Integration of hepatitis B virus: analysis of unoccupied sites

Hepatitis B virus (HBV) sequences integrated in the PLC/PRF/5 cell line (Alexander cells), which was derived from a human primary liver carcinoma, were previously extensively studied. Here we describe the analysis of the unoccupied sites of two linearly integrated forms of HBV DNA, AL-14 and AL-26, that were characterized previously. No major cellular DNA rearrangements were seen at the integration sites except for small deletions of host sequences: 2 kilobases of DNA in AL-14 and 17 base pairs (bp) in AL-26. The unoccupied site of AL-26 was found to be missing 182 bp, which previously mapped next to the right end of the integration sites of several independent clones. These were believed to be of cellular origin, but we show here that these 182 bp are in fact from unusual HBV sequences. Surprisingly, a region of this newly detected HBV DNA sequence is more homologous to that of woodchuck HBV DNA. Our analysis shows that the normal counterparts of both AL-14 and AL-26 contain minisatellite-like repetitive sequences. Based on the data presented here and our previous finding of HBV DNA integration at satellite sequences, we propose that genomic simple repetitive sequences are hot spots for HBV DNA integration.

[1]  W. Rutter,et al.  Integration of hepatitis B virus DNA in chromosome-specific satellite sequences , 1986, Journal of virology.

[2]  M. Permutt,et al.  Evidence for increased recombination near the human insulin gene: implication for disease association studies. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Swee Lay Thein,et al.  Hypervariable ‘minisatellite’ regions in human DNA , 1985, Nature.

[4]  W. Rutter,et al.  Sequence of hepatitis B virus DNA incorporated into the genome of a human hepatoma cell line , 1985, Journal of virology.

[5]  P. Hofschneider,et al.  Co‐transcribed 3′ host sequences augment expression of integrated hepatitis B virus DNA. , 1985, The EMBO journal.

[6]  W. Rutter,et al.  Hybrid hepatitis B virus-host transcripts in a human hepatoma cell. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[7]  D. Melton,et al.  Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. , 1984, Nucleic acids research.

[8]  P. Hofschneider,et al.  Amplification and rearrangement in hepatoma cell DNA associated with integrated hepatitis B virus DNA. , 1984, The EMBO journal.

[9]  M. Botchan,et al.  DNA sequence studies of simian virus 40 chromosomal excision and integration in rat cells. , 1984, Journal of molecular biology.

[10]  P. Hofschneider,et al.  The genetic organization of integrated hepatitis B virus DNA in the human hepatoma cell line PLC/PRF/5. , 1984, Nucleic acids research.

[11]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[12]  Gerald R. Smith.  Chi hotspots of generalized recombination , 1983, Cell.

[13]  S. Goodbourn,et al.  Molecular basis of length polymorphism in the human zeta-globin gene complex. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[14]  S. King,et al.  Role of short regions of homology in intermolecular illegitimate recombination events. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[15]  T. Maniatis,et al.  The structure of the human zeta-globin gene and a closely linked, nearly identical pseudogene , 1982, Cell.

[16]  W. Rutter,et al.  The highly polymorphic region near the human insulin gene is composed of simple tandemly repeating sequences , 1982, Nature.

[17]  F. Galibert,et al.  Nucleotide sequence of a cloned woodchuck hepatitis virus genome: comparison with the hepatitis B virus sequence , 1982, Journal of virology.

[18]  P. Hofschneider,et al.  Detection of hepatitis B virus-specific DNA in the genomes of human hepatocellular carcinoma and liver cirrhosis tissues. , 1981, The Journal of general virology.

[19]  W. Rutter,et al.  THE NUCLEOTIDE SEQUENCE OF THE HEPATITIS B VIRAL GENOME AND THE IDENTIFICATION OF THE MAJOR VIRAL GENES , 1980 .

[20]  R. Hardison,et al.  The isolation of structural genes from libraries of eucaryotic DNA , 1978, Cell.

[21]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[22]  P Berg,et al.  Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. , 1977, Journal of molecular biology.

[23]  W. D. Benton,et al.  Screening lambdagt recombinant clones by hybridization to single plaques in situ. , 1977, Science.

[24]  E. Southern.  Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.