Nucleic Acids Research, 1993, Vol. 21, No. 15 3507-3511
© 1993
MOLECULAR BIOLOGY |
Characterization of a DNA binding domain in the C-terminus of HIV-1 integrase by deletion mutagenesis
Division of Viral Products, Center for Biologics Evaluation and Research FDA, Bethesda, MD 20892, USA
*To whom correspondence should be addressed
Received April 6, 1993. Revised June 14, 1993. Accepted June 14, 1993.
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.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  H. Ebina, A. G. Chatterjee, R. L. Judson, and H. L. Levin The GP(Y/F) Domain of TF1 Integrase Multimerizes when Present in a Fragment, and Substitutions in This Domain Reduce Enzymatic Activity of the Full-length Protein J. Biol. Chem., June 6, 2008; 283(23): 15965 - 15974. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Ao, G. Huang, H. Yao, Z. Xu, M. Labine, A. W. Cochrane, and X. Yao Interaction of Human Immunodeficiency Virus Type 1 Integrase with Cellular Nuclear Import Receptor Importin 7 and Its Impact on Viral Replication J. Biol. Chem., May 4, 2007; 282(18): 13456 - 13467. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Topper, Y. Luo, M. Zhadina, K. Mohammed, L. Smith, and M. A. Muesing Posttranslational Acetylation of the Human Immunodeficiency Virus Type 1 Integrase Carboxyl-Terminal Domain Is Dispensable for Viral Replication J. Virol., March 15, 2007; 81(6): 3012 - 3017. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Lu, H. Z. Ghory, and A. Engelman Genetic Analyses of Conserved Residues in the Carboxyl-Terminal Domain of Human Immunodeficiency Virus Type 1 Integrase J. Virol., August 15, 2005; 79(16): 10356 - 10368. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Brigo, K. W. Lee, G. Iurcu Mustata, and J. M. Briggs Comparison of Multiple Molecular Dynamics Trajectories Calculated for the Drug-Resistant HIV-1 Integrase T66I/M154I Catalytic Domain Biophys. J., May 1, 2005; 88(5): 3072 - 3082. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Tan, K. Zhu, D. J. Segal, C. F. Barbas III, and S. A. Chow Fusion Proteins Consisting of Human Immunodeficiency Virus Type 1 Integrase and the Designed Polydactyl Zinc Finger Protein E2C Direct Integration of Viral DNA into Specific Sites J. Virol., February 1, 2004; 78(3): 1301 - 1313. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Devroe, A. Engelman, and P. A. Silver Intracellular transport of human immunodeficiency virus type 1 integrase J. Cell Sci., November 1, 2003; 116(21): 4401 - 4408. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. S. Appa, C.-G. Shin, P. Lee, and S. A. Chow Role of the Nonspecific DNA-binding Region and alpha Helices within the Core Domain of Retroviral Integrase in Selecting Target DNA Sites for Integration J. Biol. Chem., November 30, 2001; 276(49): 45848 - 45855. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. L. Holmes-Son and S. A. Chow Integrase-LexA Fusion Proteins Incorporated into Human Immunodeficiency Virus Type 1 That Contains a Catalytically Inactive Integrase Gene Are Functional To Mediate Integration J. Virol., December 15, 2000; 74(24): 11548 - 11556. [Abstract] [Full Text]
|
|
|
|
|
|
T. Ishikawa, N. Okui, N. Kobayashi, R. Sakuma, T. Kitamura, and Y. Kitamura Monoclonal Antibodies against the Minimal DNA-Binding Domain in the Carboxyl-Terminal Region of Human Immunodeficiency Virus Type 1 Integrase J. Virol., May 1, 1999; 73(5): 4475 - 4480. [Abstract] [Full Text]
|
|
|
|
|
|
F. M. I. van den Ent, A. Vos, and R. H. A. Plasterk Dissecting the Role of the N-Terminal Domain of Human Immunodeficiency Virus Integrase by trans-Complementation Analysis J. Virol., April 1, 1999; 73(4): 3176 - 3183. [Abstract] [Full Text]
|
|
|
|
|
|
F. Yang, O. Leon, N. J. Greenfield, and M. J. Roth Functional Interactions of the HHCC Domain of Moloney Murine Leukemia Virus Integrase Revealed by Nonoverlapping Complementation and Zinc-Dependent Dimerization J. Virol., March 1, 1999; 73(3): 1809 - 1817. [Abstract] [Full Text]
|
|
|
|
|
|
M. H. Nymark-McMahon and S. B. Sandmeyer Mutations in Nonconserved Domains of Ty3 Integrase Affect Multiple Stages of the Ty3 Life Cycle J. Virol., January 1, 1999; 73(1): 453 - 465. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Gaur and A. D. Leavitt Mutations in the Human Immunodeficiency Virus Type 1 Integrase D,D(35)E Motif Do Not Eliminate Provirus Formation J. Virol., June 1, 1998; 72(6): 4678 - 4685. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. P. Lutzke and R. H. A. Plasterk Structure-Based Mutational Analysis of the C-Terminal DNA-Binding Domain of Human Immunodeficiency Virus Type 1 Integrase: Critical Residues for Protein Oligomerization and DNA Binding J. Virol., June 1, 1998; 72(6): 4841 - 4848. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. M. I. van den Ent, A. Vos, and R. H. A. Plasterk Mutational Scan of the Human Immunodeficiency Virus Type 2 Integrase Protein J. Virol., May 1, 1998; 72(5): 3916 - 3924. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Dildine, J. Respess, D. Jolly, and S. B. Sandmeyer A Chimeric Ty3/Moloney Murine Leukemia Virus Integrase Protein Is Active In Vivo J. Virol., May 1, 1998; 72(5): 4297 - 4307. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Katzman and M. Sudol Mapping Viral DNA Specificity to the Central Region of Integrase by Using Functional Human Immunodeficiency Virus Type 1/Visna Virus Chimeric Proteins J. Virol., March 1, 1998; 72(3): 1744 - 1753. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Acel, B. E. Udashkin, M. A. Wainberg, and E. A. Faust Efficient Gap Repair Catalyzed In Vitro by an Intrinsic DNA Polymerase Activity of Human Immunodeficiency Virus Type 1 Integrase J. Virol., March 1, 1998; 72(3): 2062 - 2071. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. A. Donzella, O. Leon, and M. J. Roth Implication of a Central Cysteine Residue and the HHCC Domain of Moloney Murine Leukemia Virus Integrase Protein in Functional Multimerization J. Virol., February 1, 1998; 72(2): 1691 - 1698. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Asante-Appiah and A. M. Skalka A Metal-induced Conformational Change and Activation of HIV-1 Integrase J. Biol. Chem., June 27, 1997; 272(26): 16196 - 16205. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Zheng, T. M. Jenkins, and R. Craigie Zinc folds the N-terminal domain of HIV-1 integrase, promotes multimerization, and enhances catalytic activity PNAS, November 26, 1996; 93(24): 13659 - 13664. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Mazumder, N. Neamati, A. A. Pilon, S. Sunder, and Y. Pommier Chemical Trapping of Ternary Complexes of Human Immunodeficiency Virus Type 1Integrase, Divalent Metal, and DNA Substrates Containing an Abasic Site. IMPLICATIONS FOR THE ROLE OF LYSINE 136IN DNA BINDING J. Biol. Chem., November 1, 1996; 271(44): 27330 - 27338. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. D. Andrake and A. M. Skalka Retroviral Integrase, Putting the Pieces Together J. Biol. Chem., August 16, 1996; 271(33): 19633 - 19636. [Full Text] [PDF]
|
|
|
|
|
|
M. D. Andrake and A. M. Skalka Multimerization Determinants Reside in Both the Catalytic Core and C Terminus of Avian Sarcoma Virus Integrase J. Biol. Chem., December 8, 1995; 270(49): 29299 - 29306. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Pahl and R. M. Flügel Characterization of the Human Spuma Retrovirus Integrase by Site-directed Mutagenesis, by Complementation Analysis, and by Swapping the Zinc Finger Domain of HIV-1 J. Biol. Chem., February 17, 1995; 270(7): 2957 - 2966. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F Dyda, A. Hickman, T. Jenkins, A Engelman, R Craigie, and D. Davies Crystal structure of the catalytic domain of HIV-1 integrase: similarity to other polynucleotidyl transferases Science, December 23, 1994; 266(5193): 1981 - 1986. [Abstract] [PDF]
|
|
|
|
|
|
N. Jing, C. Marchand, J. Liu, R. Mitra, M. E. Hogan, and Y. Pommier Mechanism of Inhibition of HIV-1 Integrase by G-tetrad-forming Oligonucleotides in Vitro J. Biol. Chem., July 7, 2000; 275(28): 21460 - 21467. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Katz, P. DiCandeloro, G. Kukolj, and A. M. Skalka Role of DNA End Distortion in Catalysis by Avian Sarcoma Virus Integrase J. Biol. Chem., August 31, 2001; 276(36): 34213 - 34220. [Abstract] [Full Text] [PDF]
|
|