Subcellular localization of avian sarcoma virus and human immunodeficiency virus type 1 integrases

The composition and subcellular trafficking of subviral preintegration complexes are reported to vary among the different retroviruses. The process by which the avian sarcoma virus (ASV) preintegration complex gains access to target chromatin remains unknown. Here we report that ASV integrase (IN) expressed as a fusion to beta-galactosidase accumulates in the nuclei of transfected COS-1 cells. In contrast, human immunodeficiency type 1 (HIV-1) IN-beta-galactosidase fusions expressed similarly are predominantly cytoplasmic. To identify the region of ASV IN that specifies nuclear localization, various subdomains of the protein were expressed as beta-galactosidase fusions and their subcellular locations were assessed cytochemically and by indirect immunofluorescence. These analyses showed that the ASV IN protein possesses a functional nuclear localization signal that spans amino acids 206 to 235 and displays limited homology with known nuclear transport signals.

[1]  A. Skalka,et al.  Retroviral Integrase, Putting the Pieces Together* , 1996, The Journal of Biological Chemistry.

[2]  A. Kingsman,et al.  Conserved sequences in the carboxyl terminus of integrase that are essential for human immunodeficiency virus type 1 replication , 1996, Journal of virology.

[3]  A. Skalka,et al.  Multimerization Determinants Reside in Both the Catalytic Core and C Terminus of Avian Sarcoma Virus Integrase (*) , 1995, The Journal of Biological Chemistry.

[4]  R. Knoblauch,et al.  Mutational analysis of cell cycle arrest, nuclear localization and virion packaging of human immunodeficiency virus type 1 Vpr , 1995, Journal of virology.

[5]  M. Jaskólski,et al.  High-resolution structure of the catalytic domain of avian sarcoma virus integrase. , 1995, Journal of molecular biology.

[6]  Rolf Boelens,et al.  The DNA-binding domain of HIV-1 integrase has an SH3-like fold , 1995, Nature Structural Biology.

[7]  A M Gronenborn,et al.  Solution structure of the DNA binding domain of HIV-1 integrase. , 1995, Biochemistry.

[8]  D. Trono,et al.  HIV-1 infection of nondividing cells: C-terminal tyrosine phosphorylation of the viral matrix protein is a key regulator , 1995, Cell.

[9]  A. Engelman,et al.  Crystal structure of the catalytic domain of HIV-1 integrase: similarity to other polynucleotidyl transferases. , 1994, Science.

[10]  A. Skalka,et al.  Monoclonal antibodies against HIV type 1 integrase: clues to molecular structure. , 1994, AIDS research and human retroviruses.

[11]  D. Trono,et al.  The nuclear localization signal of the matrix protein of human immunodeficiency virus type 1 allows the establishment of infection in macrophages and quiescent T lymphocytes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[12]  M. Emerman,et al.  The Vpr protein of human immunodeficiency virus type 1 influences nuclear localization of viral nucleic acids in nondividing host cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. Hammarskjöld,et al.  Human immunodeficiency virus env expression becomes Rev-independent if the env region is not defined as an intron , 1994, Journal of virology.

[14]  M. Emerman,et al.  Passage through mitosis is required for oncoretroviruses but not for the human immunodeficiency virus , 1994, Journal of virology.

[15]  M. Emerman,et al.  A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells , 1993, Nature.

[16]  P. Brown,et al.  Integration of murine leukemia virus DNA depends on mitosis. , 1993, The EMBO journal.

[17]  D. Grandgenett,et al.  Characterization of a stable eukaryotic cell line expressing the Rous sarcoma virus integrase. , 1992, Virology.

[18]  M. Emerman,et al.  Human immunodeficiency virus infection of cells arrested in the cell cycle. , 1992, The EMBO journal.

[19]  M. Bukrinsky,et al.  Active nuclear import of human immunodeficiency virus type 1 preintegration complexes. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[20]  A. Skalka,et al.  Residues critical for retroviral integrative recombination in a region that is highly conserved among retroviral/retrotransposon integrases and bacterial insertion sequence transposases , 1992, Molecular and cellular biology.

[21]  A. Skalka,et al.  Identification and characterization of intragenic sequences which repress human immunodeficiency virus structural gene expression , 1991, Journal of virology.

[22]  W. Haseltine,et al.  Determination of viral proteins present in the human immunodeficiency virus type 1 preintegration complex , 1991, Journal of virology.

[23]  A. Skalka,et al.  Retroviral integrase domains: DNA binding and the recognition of LTR sequences. , 1991, Nucleic acids research.

[24]  R. Laskey,et al.  Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: Identification of a class of bipartite nuclear targeting sequence , 1991, Cell.

[25]  P. Silver.  How proteins enter the nucleus , 1991, Cell.

[26]  J. Coffin,et al.  Efficient autointegration of avian retrovirus DNA in vitro , 1990, Journal of virology.

[27]  E. Fenyö,et al.  Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1 , 1990, Journal of virology.

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

[29]  T. Copeland,et al.  rev protein of human immunodeficiency virus type 1 affects the stability and transport of the viral mRNA. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[30]  A. Skalka,et al.  Avian sarcoma-leukosis virus pol-endo proteins expressed independently in mammalian cells accumulate in the nucleus but can be directed to other cellular compartments , 1988, Journal of virology.

[31]  William D. Richardson,et al.  A short amino acid sequence able to specify nuclear location , 1984, Cell.

[32]  E. H. Humphries,et al.  Rous sarcoma virus infection of synchronized cells establishes provirus integration during S-phase DNA synthesis prior to cellular division. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Coffin,et al.  Rate of virus-specific RNA synthesis in synchronized chicken embryo fibroblasts infected with avian leukosis virus , 1976, Journal of virology.

[34]  H. Temin,et al.  Requirement for Cell Division for Initiation of Transcription of Rous Sarcoma Virus RNA , 1974, Journal of virology.

[35]  A. van der Eb,et al.  A new technique for the assay of infectivity of human adenovirus 5 DNA. , 1973, Virology.

[36]  H. Temin,et al.  Studies on carcinogenesis by avian sarcoma viruses: VIII. Glycolysis and cell multiplication , 1968 .

[37]  H. Temin.  Studies on carcinogenesis by avian sarcoma viruses. V. Requirement for new DNA synthesis and for cell division , 1967 .

[38]  M. Stevenson.  Portals of entry: uncovering HIV nuclear transport pathways. , 1996, Trends in cell biology.

[39]  A. Skalka,et al.  The retroviral enzymes. , 1994, Annual review of biochemistry.

[40]  T. Boulikas,et al.  Nuclear localization signals (NLS). , 1993, Critical reviews in eukaryotic gene expression.

[41]  S. Goff,et al.  Genetics of retroviral integration. , 1992, Annual review of genetics.

[42]  R. Laskey,et al.  Nuclear targeting sequences--a consensus? , 1991, Trends in biochemical sciences.

[43]  Ajit Kumar.  Advances in Molecular Biology and Targeted Treatment for AIDS , 1991, GWUMC Department of Biochemistry Annual Spring Symposia.

[44]  A. Skalka,et al.  Analyses of HIV Integration Components , 1991 .

[45]  A. Martinez-Arias,et al.  23 – β-Galactosidase Gene Fusions for Analyzing Gene Expression in Escherichia coli and Yeast , 1989 .

[46]  A. Martinez-Arias,et al.  Beta-galactosidase gene fusions for analyzing gene expression in escherichia coli and yeast. , 1983, Methods in enzymology.