Home LiteratureArticle Details
PMID: 8107210 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Activities of the feline immunodeficiency virus integrase protein produced in Escherichia coli.

Journal of virology ·Vol. 68 ·No. 3 ·1994-03-00 ·Pages 1468-74

Vink C, van der Linden KH, Plasterk RH

Abstract

Retroviral DNA integration requires the activity of at least one viral protein, the integrase (IN) protein. We cloned and expressed the integrase gene of feline immunodeficiency virus (FIV) in Escherichia coli as a fusion to the malE gene and purified the IN fusion protein by affinity chromatography. The protein is active in site-specific cleavage of the viral DNA ends, DNA strand transfer, and disintegration. FIV IN has a relaxed viral DNA substrate requirement: it cleaves and integrates FIV DNA termini, human immunodeficiency virus DNA ends, and Moloney murine leukemia virus DNA ends with high efficiencies. In the cleavage reaction, IN exposes a specific phosphodiester bond near the viral DNA end to nucleophilic attack. In vitro, either H2O, glycerol, or the 3' OH group of the viral DNA terminus can serve as nucleophile in this reaction. We found that FIV IN preferentially uses the 3' OH ends of the viral DNA as nucleophile, whereas HIV IN protein preferentially uses H2O and glycerol as nucleophiles.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular DNA Nucleotidyltransferases/genetics,isolation & purification,metabolism DNA Transposable Elements DNA-Binding Proteins/metabolism Escherichia coli/genetics Immunodeficiency Virus, Feline/enzymology Integrases Molecular Sequence Data Recombinant Fusion Proteins/isolation & purification,metabolism Sequence Homology, Amino Acid Substrate Specificity Virus Integration/physiology
Chemicals
DNA Transposable Elements DNA-Binding Proteins Recombinant Fusion Proteins DNA Nucleotidyltransferases Integrases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Vink C
Division of Molecular Biology, The Netherlands Cancer Institute, Amsterdam.
van der Linden K H
Plasterk R H
References (48)
48 references, click to expand
  1. Cloning of HTLV-4 and its relation to simian and human immunodeficiency viruses.
    Nature. 1987 Apr 9-15;326(6113):610-3 PMID: 3104797
  2. The human immunodeficiency virus integrase protein.
    Trends Genet. 1993 Dec;9(12):433-8 PMID: 8122311
  3. A simple salting out procedure for extracting DNA from human nucleated cells.
    Nucleic Acids Res. 1988 Feb 11;16(3):1215 PMID: 3344216
  4. Gene product of Moloney murine leukemia virus required for proviral integration is a DNA-binding protein.
    J Mol Biol. 1988 Sep 5;203(1):131-9 PMID: 3054118
  5. Integration of mini-retroviral DNA: a cell-free reaction for biochemical analysis of retroviral integration.
    Proc Natl Acad Sci U S A. 1989 May;86(9):3065-9 PMID: 2524066
  6. Nucleotide sequence and genomic organization of feline immunodeficiency virus.
    Proc Natl Acad Sci U S A. 1989 Aug;86(15):5743-7 PMID: 2762293
  7. The avian retroviral integration protein cleaves the terminal sequences of linear viral DNA at the in vivo sites of integration.
    J Virol. 1989 Dec;63(12):5319-27 PMID: 2555556
  8. Unraveling retrovirus integration.
    Cell. 1990 Jan 12;60(1):3-4 PMID: 2403842
  9. Nucleotide sequence and genome organization of biologically active proviruses of the bovine immunodeficiency-like virus.
    Virology. 1990 Apr;175(2):391-409 PMID: 2183467
  10. Human immunodeficiency virus integration protein expressed in Escherichia coli possesses selective DNA cleaving activity.
    Proc Natl Acad Sci U S A. 1990 Jul;87(13):5119-23 PMID: 2164223
  11. The IN protein of Moloney murine leukemia virus processes the viral DNA ends and accomplishes their integration in vitro.
    Cell. 1990 Aug 24;62(4):829-37 PMID: 2167180
  12. Integration of retroviral DNA.
    Curr Top Microbiol Immunol. 1990;157:19-48 PMID: 2203610
  13. Integration of human immunodeficiency virus types 1 and 2 DNA in vitro by cytoplasmic extracts of Moloney murine leukemia virus-infected mouse NIH 3T3 cells.
    J Virol. 1990 Oct;64(10):5219-22 PMID: 2398544
  14. The avian retroviral IN protein is both necessary and sufficient for integrative recombination in vitro.
    Cell. 1990 Oct 5;63(1):87-95 PMID: 2170022
  15. Defining nucleic acid-binding properties of avian retrovirus integrase by deletion analysis.
    J Virol. 1991 Mar;65(3):1160-7 PMID: 1847445
  16. Activities of human immunodeficiency virus (HIV) integration protein in vitro: specific cleavage and integration of HIV DNA.
    Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1339-43 PMID: 1847518
  17. Functional similarities between retroviruses and the IS3 family of bacterial insertion sequences?
    Mol Microbiol. 1990 Oct;4(10):1771-7 PMID: 1963920
  18. Retroviral integrase domains: DNA binding and the recognition of LTR sequences.
    Nucleic Acids Res. 1991 Feb 25;19(4):851-60 PMID: 1850126
  19. DNA binding properties of the integrase proteins of human immunodeficiency viruses types 1 and 2.
    Nucleic Acids Res. 1991 Jul 25;19(14):3821-7 PMID: 1861975
  20. Human immunodeficiency virus integrase protein requires a subterminal position of its viral DNA recognition sequence for efficient cleavage.
    J Virol. 1991 Sep;65(9):4636-44 PMID: 1870194
  21. Substrate specificity of recombinant human immunodeficiency virus integrase protein.
    J Virol. 1991 Oct;65(10):5624-30 PMID: 1895409
  22. Development of IL-2-independent feline lymphoid cell lines chronically infected with feline immunodeficiency virus: importance for diagnostic reagents and vaccines.
    Intervirology. 1991;32(6):361-75 PMID: 1657826
  23. Site-specific hydrolysis and alcoholysis of human immunodeficiency virus DNA termini mediated by the viral integrase protein.
    Nucleic Acids Res. 1991 Dec 25;19(24):6691-8 PMID: 1662361
  24. Reversal of integration and DNA splicing mediated by integrase of human immunodeficiency virus.
    Science. 1992 Feb 7;255(5045):723-6 PMID: 1738845
  25. Both substrate and target oligonucleotide sequences affect in vitro integration mediated by human immunodeficiency virus type 1 integrase protein produced in Saccharomyces cerevisiae.
    J Virol. 1992 Apr;66(4):2359-68 PMID: 1548767
  26. Residues critical for retroviral integrative recombination in a region that is highly conserved among retroviral/retrotransposon integrases and bacterial insertion sequence transposases.
    Mol Cell Biol. 1992 May;12(5):2331-8 PMID: 1314954
  27. Human immunodeficiency virus type 1 integration protein: DNA sequence requirements for cleaving and joining reactions.
    J Virol. 1992 Jun;66(6):3593-601 PMID: 1374809
  28. Nucleotide sequence and transcriptional analysis of molecular clones of CAEV which generate infectious virus.
    Virology. 1990 Nov;179(1):347-64 PMID: 2171210
  29. The N-terminal region of HIV-1 integrase is required for integration activity, but not for DNA-binding.
    Biochem Biophys Res Commun. 1992 Jun 30;185(3):874-80 PMID: 1627142
  30. Retroviral integrase functions as a multimer and can turn over catalytically.
    J Biol Chem. 1992 Aug 15;267(23):16037-40 PMID: 1322888
  31. Hotspots and warm spots: integration specificity of retroelements.
    Trends Genet. 1992 Jun;8(6):187-90 PMID: 1323152
  32. Polynucleotidyl transfer reactions in transpositional DNA recombination.
    J Biol Chem. 1992 Oct 25;267(30):21273-6 PMID: 1383220
  33. Mutational analysis of the integrase protein of human immunodeficiency virus type 2.
    Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9598-602 PMID: 1409671
  34. Characterization of human immunodeficiency virus type 1 integrase expressed in Escherichia coli and analysis of variants with amino-terminal mutations.
    J Virol. 1993 Jan;67(1):425-37 PMID: 8416376
  35. Genetics of retroviral integration.
    Annu Rev Genet. 1992;26:527-44 PMID: 1482125
  36. Identification of proteolytic processing sites within the Gag and Pol polyproteins of feline immunodeficiency virus.
    J Virol. 1993 Apr;67(4):1869-76 PMID: 8383214
  37. Identification of the catalytic and DNA-binding region of the human immunodeficiency virus type I integrase protein.
    Nucleic Acids Res. 1993 Mar 25;21(6):1419-25 PMID: 8464733
  38. Domains of the integrase protein of human immunodeficiency virus type 1 responsible for polynucleotidyl transfer and zinc binding.
    Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3428-32 PMID: 8386373
  39. A 32,000-dalton nucleic acid-binding protein from avian retravirus cores possesses DNA endonuclease activity.
    Virology. 1978 Aug;89(1):119-32 PMID: 210568
  40. Nucleotide sequence of Moloney murine leukaemia virus.
    Nature. 1981 Oct 15-21;293(5833):543-8 PMID: 6169994
  41. Nucleotide sequence of Rous sarcoma virus.
    Cell. 1983 Mar;32(3):853-69 PMID: 6299578
  42. Purification of genomic sequences from bacteriophage libraries by recombination and selection in vivo.
    Nucleic Acids Res. 1983 Apr 25;11(8):2427-45 PMID: 6856467
  43. Nucleotide sequence of the AIDS virus, LAV.
    Cell. 1985 Jan;40(1):9-17 PMID: 2981635
  44. Isolation of a T-lymphotropic virus from domestic cats with an immunodeficiency-like syndrome.
    Science. 1987 Feb 13;235(4790):790-3 PMID: 3643650
  45. Complementation between HIV integrase proteins mutated in different domains.
    EMBO J. 1993 Aug;12(8):3261-7 PMID: 8344263
  46. Identification of discrete functional domains of HIV-1 integrase and their organization within an active multimeric complex.
    EMBO J. 1993 Aug;12(8):3269-75 PMID: 8344264
  47. Identification of amino acids in HIV-2 integrase involved in site-specific hydrolysis and alcoholysis of viral DNA termini.
    Nucleic Acids Res. 1993 Jul 25;21(15):3373-7 PMID: 8346016
  48. Strong sequence conservation among horizontally transmissible, minimally pathogenic feline leukemia viruses.
    J Virol. 1988 Mar;62(3):722-31 PMID: 2828667
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.68.3.1468-1474.1994
Published
1994-03-00
Pages
1468-74
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC236602
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.68.3.1468-1474.1994 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2