Search tips
Search criteria 


Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. 1996 March; 70(3): 1580–1587.
PMCID: PMC189980

Monoclonal antibodies against human immunodeficiency virus type 1 integrase: epitope mapping and differential effects on integrase activities in vitro.


Human immunodeficiency virus type 1 (HIV-1) integrase (IN) catalyzes the integration of viral DNA into the host chromosome, an essential step in retroviral replication. As a tool to study the structure and function of this enzyme, monoclonal antibodies (MAbs) against HIV-1 IN were produced. Epitope mapping demonstrated that the 17 MAbs obtained could be divided into seven different groups, and the selection of MAbs representing these groups were tested for their effect on in vitro activities of IN. Four groups of MAbs recognized epitopes within the region of amino acids (aa) 1 to 16, 17 to 38, or 42 to 55 in and around the conserved HHCC motif near the N terminus of IN. MAbs binding to these epitopes inhibited end processing and DNA joining and either stimulated or had little effect on disintegration and reintegration activities of IN. Two MAbs binding to epitopes within the region of aa 56 to 102 in the central core or aa 186 to 250 in the C-terminal half of the protein showed only minor effects on the in vitro activities of IN. Three Mabs which recognized on epitope within the region of aa262 to 271 of HIV-1 IN cross-reacted with HIV-2 IN. MAbs binding to this epitope clearly inhibited end processing and DNA joining and stimulated or had little effect on disintegration. In contrast to the N-terminal-specific MAbs, these C-terminal-specific MAbs abolished reintegration activity of IN.

Full Text

The Full Text of this article is available as a PDF (357K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allen P, Worland S, Gold L. Isolation of high-affinity RNA ligands to HIV-1 integrase from a random pool. Virology. 1995 Jun 1;209(2):327–336. [PubMed]
  • Bizub-Bender D, Kulkosky J, Skalka AM. Monoclonal antibodies against HIV type 1 integrase: clues to molecular structure. AIDS Res Hum Retroviruses. 1994 Sep;10(9):1105–1115. [PubMed]
  • Björling E, Utter G, Stålhandske P, Norrby E, Chiodi F. Identification of a uniquely immunodominant, cross-reacting site in the human immunodeficiency virus endonuclease protein. J Virol. 1991 Aug;65(8):4543–4546. [PMC free article] [PubMed]
  • Brown PO. Integration of retroviral DNA. Curr Top Microbiol Immunol. 1990;157:19–48. [PubMed]
  • Brown PO, Bowerman B, Varmus HE, Bishop JM. Retroviral integration: structure of the initial covalent product and its precursor, and a role for the viral IN protein. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2525–2529. [PubMed]
  • Burke CJ, Sanyal G, Bruner MW, Ryan JA, LaFemina RL, Robbins HL, Zeft AS, Middaugh CR, Cordingley MG. Structural implications of spectroscopic characterization of a putative zinc finger peptide from HIV-1 integrase. J Biol Chem. 1992 May 15;267(14):9639–9644. [PubMed]
  • Bushman FD, Craigie R. 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):1339–1343. [PubMed]
  • Bushman FD, Engelman A, Palmer I, Wingfield P, Craigie R. 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):3428–3432. [PubMed]
  • Bushman FD, Fujiwara T, Craigie R. Retroviral DNA integration directed by HIV integration protein in vitro. Science. 1990 Sep 28;249(4976):1555–1558. [PubMed]
  • Chow SA, Vincent KA, Ellison V, Brown PO. Reversal of integration and DNA splicing mediated by integrase of human immunodeficiency virus. Science. 1992 Feb 7;255(5045):723–726. [PubMed]
  • Drelich M, Wilhelm R, Mous J. Identification of amino acid residues critical for endonuclease and integration activities of HIV-1 IN protein in vitro. Virology. 1992 Jun;188(2):459–468. [PubMed]
  • Ellison V, Brown PO. A stable complex between integrase and viral DNA ends mediates human immunodeficiency virus integration in vitro. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7316–7320. [PubMed]
  • Ellison V, Gerton J, Vincent KA, Brown PO. An essential interaction between distinct domains of HIV-1 integrase mediates assembly of the active multimer. J Biol Chem. 1995 Feb 17;270(7):3320–3326. [PubMed]
  • Engelman A, Craigie R. Identification of conserved amino acid residues critical for human immunodeficiency virus type 1 integrase function in vitro. J Virol. 1992 Nov;66(11):6361–6369. [PMC free article] [PubMed]
  • Engelman A, Hickman AB, Craigie R. The core and carboxyl-terminal domains of the integrase protein of human immunodeficiency virus type 1 each contribute to nonspecific DNA binding. J Virol. 1994 Sep;68(9):5911–5917. [PMC free article] [PubMed]
  • Fujiwara T, Mizuuchi K. Retroviral DNA integration: structure of an integration intermediate. Cell. 1988 Aug 12;54(4):497–504. [PubMed]
  • Goff SP. Genetics of retroviral integration. Annu Rev Genet. 1992;26:527–544. [PubMed]
  • Katz RA, Merkel G, Kulkosky J, Leis J, Skalka AM. The avian retroviral IN protein is both necessary and sufficient for integrative recombination in vitro. Cell. 1990 Oct 5;63(1):87–95. [PubMed]
  • Khan E, Mack JP, Katz RA, Kulkosky J, Skalka AM. Retroviral integrase domains: DNA binding and the recognition of LTR sequences. Nucleic Acids Res. 1991 Feb 25;19(4):851–860. [PMC free article] [PubMed]
  • Kulkosky J, Jones KS, Katz RA, Mack JP, Skalka AM. 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):2331–2338. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lutzke RA, Vink C, Plasterk RH. Characterization of the minimal DNA-binding domain of the HIV integrase protein. Nucleic Acids Res. 1994 Oct 11;22(20):4125–4131. [PMC free article] [PubMed]
  • Schauer M, Billich A. 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):874–880. [PubMed]
  • Sherman PA, Fyfe JA. 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):5119–5123. [PubMed]
  • Szilvay AM, Nornes S, Haugan IR, Olsen L, Prasad VR, Endresen C, Goff SP, Helland DE. Epitope mapping of HIV-1 reverse transcriptase with monoclonal antibodies that inhibit polymerase and RNase H activities. J Acquir Immune Defic Syndr. 1992;5(7):647–657. [PubMed]
  • Tanese N, Prasad VR, Goff SP. Structural requirements for bacterial expression of stable, enzymatically active fusion proteins containing the human immunodeficiency virus reverse transcriptase. DNA. 1988 Jul-Aug;7(6):407–416. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Vincent KA, Ellison V, Chow SA, Brown PO. 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):425–437. [PMC free article] [PubMed]
  • Vinga-Martins C, Schneider T, Werno A, Roenspeck W, Pauli G, Mueller-Lantzsch N. Mapping of immunodominant epitopes of the HIV-1 and HIV-2 integrase proteins by recombinant proteins and synthetic peptides. AIDS Res Hum Retroviruses. 1992 Jul;8(7):1301–1310. [PubMed]
  • Vink C, Oude Groeneger AM, Plasterk RH. 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):1419–1425. [PMC free article] [PubMed]
  • Whitcomb JM, Hughes SH. Retroviral reverse transcription and integration: progress and problems. Annu Rev Cell Biol. 1992;8:275–306. [PubMed]
  • Woerner AM, Marcus-Sekura CJ. Characterization of a DNA binding domain in the C-terminus of HIV-1 integrase by deletion mutagenesis. Nucleic Acids Res. 1993 Jul 25;21(15):3507–3511. [PMC free article] [PubMed]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)