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PMID: 8344264 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of discrete functional domains of HIV-1 integrase and their organization within an active multimeric complex.

The EMBO journal ·Vol. 12 ·No. 8 ·1993-08-00 ·Pages 3269-75

Engelman A, Bushman FD, Craigie R

Abstract

HIV-1 integrase protein possesses the 3' processing and DNA strand transfer activities that are required to integrate HIV DNA into a host chromosome. The N-, C-terminal and core domains of integrase are necessary for both activities in vitro. We find that certain pairs of mutant integrase proteins, which are inactive when each protein is assayed alone, can support near wild type levels of activity when both proteins are present together in the reaction mixture. This complementation implies that HIV-1 integrase functions as a multimer and has enabled us to probe the organization of the functional domains within active mixed multimers. We have identified a minimal set of functional integrase domains that are sufficient for 3' processing and DNA strand transfer and find that some domains are contributed in trans by separate monomers within the functional complex.

MeSH Terms
Base Sequence Catalysis DNA/metabolism DNA Nucleotidyltransferases/chemistry,metabolism HIV-1/enzymology Integrases Molecular Sequence Data Protein Processing, Post-Translational Retroviridae Proteins/chemistry,metabolism Virus Integration
Chemicals
Retroviridae Proteins DNA DNA Nucleotidyltransferases Integrases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Engelman A
Laboratory of Molecular Biology, National Institute of Diabetes, Digestive and Kidney Diseases, Bethesda, MD 20892.
Bushman F D
Craigie R
References (39)
39 references, click to expand
  1. Retroviral reverse transcription and integration: progress and problems.
    Annu Rev Cell Biol. 1992;8:275-306 PMID: 1282352
  2. Substrate specificity of recombinant human immunodeficiency virus integrase protein.
    J Virol. 1991 Oct;65(10):5624-30 PMID: 1895409
  3. Site-directed mutagenesis of HIV-1 integrase demonstrates differential effects on integrase functions in vitro.
    J Biol Chem. 1993 Jan 25;268(3):2113-9 PMID: 8420982
  4. 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
  5. Complementation between HIV integrase proteins mutated in different domains.
    EMBO J. 1993 Aug;12(8):3261-7 PMID: 8344263
  6. Silver staining of proteins in polyacrylamide gels.
    Anal Biochem. 1981 Nov 15;118(1):197-203 PMID: 6175245
  7. Computer analysis of retroviral pol genes: assignment of enzymatic functions to specific sequences and homologies with nonviral enzymes.
    Proc Natl Acad Sci U S A. 1986 Oct;83(20):7648-52 PMID: 2429313
  8. Correct integration of retroviral DNA in vitro.
    Cell. 1987 May 8;49(3):347-56 PMID: 3032450
  9. Retroviral DNA integration: structure of an integration intermediate.
    Cell. 1988 Aug 12;54(4):497-504 PMID: 3401925
  10. A nucleoprotein complex mediates the integration of retroviral DNA.
    Genes Dev. 1989 Apr;3(4):469-78 PMID: 2721960
  11. Human immunodeficiency virus integration in a cell-free system.
    J Virol. 1990 Jun;64(6):2711-5 PMID: 2335814
  12. Integration of human immunodeficiency virus type 1 DNA in vitro.
    Proc Natl Acad Sci U S A. 1990 Jun;87(11):4164-8 PMID: 2349226
  13. 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
  14. Retroviral DNA integration directed by HIV integration protein in vitro.
    Science. 1990 Sep 28;249(4976):1555-8 PMID: 2171144
  15. 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
  16. Determination of viral proteins present in the human immunodeficiency virus type 1 preintegration complex.
    J Virol. 1991 Apr;65(4):1910-5 PMID: 2002549
  17. DNase protection analysis of the stable synaptic complexes involved in Mu transposition.
    Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9031-5 PMID: 1656459
  18. HIV-1 DNA integration: mechanism of viral DNA cleavage and DNA strand transfer.
    Cell. 1991 Dec 20;67(6):1211-21 PMID: 1760846
  19. 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
  20. Reversal of integration and DNA splicing mediated by integrase of human immunodeficiency virus.
    Science. 1992 Feb 7;255(5045):723-6 PMID: 1738845
  21. Localization of DNA binding activity of HIV-1 integrase to the C-terminal half of the protein.
    AIDS Res Hum Retroviruses. 1992 Feb;8(2):297-304 PMID: 1540416
  22. 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
  23. Structural implications of spectroscopic characterization of a putative zinc finger peptide from HIV-1 integrase.
    J Biol Chem. 1992 May 15;267(14):9639-44 PMID: 1577801
  24. Identification of amino acid residues critical for endonuclease and integration activities of HIV-1 IN protein in vitro.
    Virology. 1992 Jun;188(2):459-68 PMID: 1585629
  25. DNA cleavage in trans by the active site tyrosine during Flp recombination: switching protein partners before exchanging strands.
    Cell. 1992 May 15;69(4):647-58 PMID: 1586945
  26. 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
  27. Assembly of the active form of the transposase-Mu DNA complex: a critical control point in Mu transposition.
    Cell. 1992 Jul 24;70(2):303-11 PMID: 1322248
  28. Retroviral integrase functions as a multimer and can turn over catalytically.
    J Biol Chem. 1992 Aug 15;267(23):16037-40 PMID: 1322888
  29. Transpositional recombination: mechanistic insights from studies of mu and other elements.
    Annu Rev Biochem. 1992;61:1011-51 PMID: 1323232
  30. Identification of conserved amino acid residues critical for human immunodeficiency virus type 1 integrase function in vitro.
    J Virol. 1992 Nov;66(11):6361-9 PMID: 1404595
  31. 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
  32. Requirement of active human immunodeficiency virus type 1 integrase enzyme for productive infection of human T-lymphoid cells.
    J Virol. 1992 Dec;66(12):7414-9 PMID: 1433523
  33. 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
  34. Epitope tagging and protein surveillance.
    Methods Enzymol. 1991;194:508-19 PMID: 1706460
  35. Retroviral integrase domains: DNA binding and the recognition of LTR sequences.
    Nucleic Acids Res. 1991 Feb 25;19(4):851-60 PMID: 1850126
  36. 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
  37. 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
  38. Retrovirus integrase: identification of a potential leucine zipper motif.
    Protein Eng. 1991 Apr;4(4):435-41 PMID: 1652750
  39. Genetics of retroviral integration.
    Annu Rev Genet. 1992;26:527-44 PMID: 1482125
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-08-00
Pages
3269-75
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413594
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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