Characterization of a DNA binding domain in the C-terminus of HIV-1 integrase by deletion mutagenesis - PubMed (original) (raw)
Characterization of a DNA binding domain in the C-terminus of HIV-1 integrase by deletion mutagenesis
A M Woerner et al. Nucleic Acids Res. 1993.
Free PMC article
Abstract
The integrase (IN) protein of human immunodeficiency virus type 1 (HIV-1) catalyzes site-specific cleavage of 2 bases from the viral long terminal repeat (LTR) sequence yet it binds DNA with little DNA sequence specificity. We have previously demonstrated that the C-terminal half of IN (amino acids 154-288) possesses a DNA binding domain. In order to further characterize this region, a series of clones expressing truncated forms of IN as N-terminal fusion proteins in E.coli were constructed and analyzed by Southwestern blotting. Proteins containing amino acids 1-263, 1-248 and 170-288 retained the ability to bind DNA, whereas a protein containing amino acids 1-180 showed no detectable DNA binding. This defines a DNA binding domain contained within amino acids 180-248. This region contains an arrangement of 9 lysine and arginine residues each separated by 2-4 amino acids (KxxxKxxxKxxxxRxxxRxxRxxxxKxxxKxxxK), spanning amino acids 211-244, which is conserved in all HIV-1 isolates. A clone expressing full-length IN with a C-terminal fusion of 16 amino acids was able to bind DNA comparably to a cloned protein with a free C-terminus, and an IN-specific monoclonal antibody which recognizes an epitope contained within amino acids 264-279 was unable to block DNA binding, supporting the evidence that a region necessary for binding lies upstream of amino acid 264.
Similar articles
- Characterization of the minimal DNA-binding domain of the HIV integrase protein.
Lutzke RA, Vink C, Plasterk RH. Lutzke RA, et al. Nucleic Acids Res. 1994 Oct 11;22(20):4125-31. doi: 10.1093/nar/22.20.4125. Nucleic Acids Res. 1994. PMID: 7937137 Free PMC article. - Localization of DNA binding activity of HIV-1 integrase to the C-terminal half of the protein.
Woerner AM, Klutch M, Levin JG, Marcus-Sekura CJ. Woerner AM, et al. AIDS Res Hum Retroviruses. 1992 Feb;8(2):297-304. doi: 10.1089/aid.1992.8.297. AIDS Res Hum Retroviruses. 1992. PMID: 1540416 - Expression of HIV-1 integrase in E. coli: immunological analysis of the recombinant protein.
Marcus-Sekura CJ, Woerner AM, Zweig M, Court DL, Levin JG, Klutch M. Marcus-Sekura CJ, et al. AIDS Res Hum Retroviruses. 1990 Dec;6(12):1399-408. doi: 10.1089/aid.1990.6.1399. AIDS Res Hum Retroviruses. 1990. PMID: 2078417 - Retroviral integrases and their cousins.
Rice P, Craigie R, Davies DR. Rice P, et al. Curr Opin Struct Biol. 1996 Feb;6(1):76-83. doi: 10.1016/s0959-440x(96)80098-4. Curr Opin Struct Biol. 1996. PMID: 8696976 Review.
Cited by
- The HIV-1 Integrase C-Terminal Domain Induces TAR RNA Structural Changes Promoting Tat Binding.
Rocchi C, Louvat C, Miele AE, Batisse J, Guillon C, Ballut L, Lener D, Negroni M, Ruff M, Gouet P, Fiorini F. Rocchi C, et al. Int J Mol Sci. 2022 Nov 8;23(22):13742. doi: 10.3390/ijms232213742. Int J Mol Sci. 2022. PMID: 36430221 Free PMC article. - Differential multimerization of Moloney murine leukemia virus integrase purified under nondenaturing conditions.
Villanueva RA, Jonsson CB, Jones J, Georgiadis MM, Roth MJ. Villanueva RA, et al. Virology. 2003 Nov 10;316(1):146-60. doi: 10.1016/s0042-6822(03)00559-2. Virology. 2003. PMID: 14599799 Free PMC article. - Characterization of the minimal DNA-binding domain of the HIV integrase protein.
Lutzke RA, Vink C, Plasterk RH. Lutzke RA, et al. Nucleic Acids Res. 1994 Oct 11;22(20):4125-31. doi: 10.1093/nar/22.20.4125. Nucleic Acids Res. 1994. PMID: 7937137 Free PMC article. - Critical contacts between HIV-1 integrase and viral DNA identified by structure-based analysis and photo-crosslinking.
Jenkins TM, Esposito D, Engelman A, Craigie R. Jenkins TM, et al. EMBO J. 1997 Nov 17;16(22):6849-59. doi: 10.1093/emboj/16.22.6849. EMBO J. 1997. PMID: 9362498 Free PMC article. - Monoclonal antibodies against the minimal DNA-binding domain in the carboxyl-terminal region of human immunodeficiency virus type 1 integrase.
Ishikawa T, Okui N, Kobayashi N, Sakuma R, Kitamura T, Kitamura Y. Ishikawa T, et al. J Virol. 1999 May;73(5):4475-80. doi: 10.1128/JVI.73.5.4475-4480.1999. J Virol. 1999. PMID: 10196350 Free PMC article.
References
- Mol Cell Biol. 1992 May;12(5):2331-8 - PubMed
- AIDS Res Hum Retroviruses. 1992 Feb;8(2):297-304 - PubMed
- Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6580-4 - PubMed
- J Biol Chem. 1992 Aug 15;267(23):16037-40 - PubMed
- Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6741-5 - PubMed
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources