Human immunodeficiency virus integration in a cell-free system

Integration of the viral genome into the nuclear DNA of a host cell plays a pivotal role in the replication of retroviruses. We have developed an in vitro method for studying the biochemistry of human immunodeficiency virus (HIV) integration by using extracts from HIV-infected cells. Analysis of the reaction products showed that HIV integration in vitro accurately reproduces the in vivo process. Integration occurred without apparent specificity for the target sequence, and the integrated provirus was directly flanked by a 5-base-pair duplication of DNA from the target site. HIV integration did not require a high-energy cofactor, and the enzymatic activities required for integration were recovered with the viral DNA when cell extracts were fractionated by gel exclusion chromatography.

[1]  T. Quinn,et al.  The international epidemiology of AIDS. , 1988, Scientific American.

[2]  P. Brown,et al.  Correct integration of retroviral DNA in vitro , 1987, Cell.

[3]  L. Lasky,et al.  Nucleic acid structure and expression of the human AIDS/lymphadenopathy retrovirus , 1985, Nature.

[4]  J. Curran,et al.  The epidemiology of AIDS in the U.S. , 1988, Scientific American.

[5]  H. Jaffe,et al.  Epidemiology of HIV infection and AIDS in the United States. , 1988, Science.

[6]  M. Reitz,et al.  In vitro generation of an HTLV-III variant by neutralizing antibody. , 1986, Journal of immunology.

[7]  J. Boeke,et al.  The DNA intermediate in yeast Ty1 element transposition copurifies with virus-like particles: Cell-free Ty1 transposition , 1988, Cell.

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

[9]  C. Lutz,et al.  Syrinx 2A: an improved lambda phage vector designed for screening DNA libraries by recombination in vivo. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[10]  A. Skalka,et al.  The avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration , 1989, Journal of virology.

[11]  B. Haynes,et al.  Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. , 1984, Science.

[12]  T. Lebien.  RNA Tumor Viruses , 1975 .

[13]  K. Mizuuchi,et al.  Retroviral DNA integration: Structure of an integration intermediate , 1988, Cell.

[14]  R. Craigie,et al.  Integration of mini-retroviral DNA: a cell-free reaction for biochemical analysis of retroviral integration. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Mark L. Pearson,et al.  Complete nucleotide sequence of the AIDS virus, HTLV-III , 1985, Nature.

[16]  P. Brown,et al.  A nucleoprotein complex mediates the integration of retroviral DNA. , 1989, Genes & development.

[17]  P. Brown,et al.  Retroviral integration: structure of the initial covalent product and its precursor, and a role for the viral IN protein. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[18]  L. Ratner,et al.  Characterization of long terminal repeat sequences of HTLV-III. , 1985, Science.