This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lutzke, R. A. P.
Right arrow Articles by Plasterk, R. H. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lutzke, R. A. P.
Right arrow Articles by Plasterk, R. H. A.

 Previous Article  |  Next Article 

J Virol, June 1998, p. 4841-4848, Vol. 72, No. 6
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

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

Ramon A. Puras Lutzkedagger and Ronald H. A. Plasterk*

Division of Molecular Biology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands

Received 3 November 1997/Accepted 3 March 1998

The C-terminal domain of human immunodeficiency virus type 1 (HIV-1) integrase (IN) is a dimer that binds to DNA in a nonspecific manner. The structure of the minimal region required for DNA binding (IN220-270) has been solved by nuclear magnetic resonance spectroscopy. The overall fold of the C-terminal domain of HIV-1 IN is similar to those of Src homology region 3 domains. Based on the structure of IN220-270, we studied the role of 15 amino acid residues potentially involved in DNA binding and oligomerization by mutational analysis. We found that two amino acid residues, arginine 262 and leucine 234, contribute to DNA binding in the context of IN220-270, as indicated by protein-DNA UV cross-link analysis. We also analyzed mutant proteins representing portions of the full-length IN protein. Amino acid substitution of residues located in the hydrophobic dimer interface, such as L241A and L242A, results in the loss of oligomerization of IN; consequently, the levels of 3' processing, DNA strand transfer, and intramolecular disintegration are strongly reduced. These results suggest that dimerization of the C-terminal domain of IN is important for correct multimerization of IN.

* Corresponding author. Mailing address: Division of Molecular Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. Phone: 31-20-5122081. Fax: 31-20-6691386. E-mail: rplas{at}nki.nl.

dagger Present address: Department of Biochemistry and Molecular Biology, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands.

J Virol, June 1998, p. 4841-4848, Vol. 72, No. 6
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Ebina, H., Chatterjee, A. G., Judson, R. L., Levin, H. L. (2008). 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. 283: 15965-15974 [Abstract] [Full Text]  
  • Pandey, K. K., Bera, S., Zahm, J., Vora, A., Stillmock, K., Hazuda, D., Grandgenett, D. P. (2007). Inhibition of Human Immunodeficiency Virus Type 1 Concerted Integration by Strand Transfer Inhibitors Which Recognize a Transient Structural Intermediate. J. Virol. 81: 12189-12199 [Abstract] [Full Text]  
  • Topper, M., Luo, Y., Zhadina, M., Mohammed, K., Smith, L., Muesing, M. A. (2007). Posttranslational Acetylation of the Human Immunodeficiency Virus Type 1 Integrase Carboxyl-Terminal Domain Is Dispensable for Viral Replication. J. Virol. 81: 3012-3017 [Abstract] [Full Text]  
  • Baranova, S., Tuzikov, F. V., Zakharova, O. D., Tuzikova, N. A., Calmels, C., Litvak, S., Tarrago-Litvak, L., Parissi, V., Nevinsky, G. A. (2007). Small-angle X-ray characterization of the nucleoprotein complexes resulting from DNA-induced oligomerization of HIV-1 integrase. Nucleic Acids Res 35: 975-987 [Abstract] [Full Text]  
  • Puglia, J., Wang, T., Smith-Snyder, C., Cote, M., Scher, M., Pelletier, J. N., John, S., Jonsson, C. B., Roth, M. J. (2006). Revealing Domain Structure through Linker-Scanning Analysis of the Murine Leukemia Virus (MuLV) RNase H and MuLV and Human Immunodeficiency Virus Type 1 Integrase Proteins. J. Virol. 80: 9497-9510 [Abstract] [Full Text]  
  • Chen, A., Weber, I. T., Harrison, R. W., Leis, J. (2006). Identification of Amino Acids in HIV-1 and Avian Sarcoma Virus Integrase Subsites Required for Specific Recognition of the Long Terminal Repeat Ends. J. Biol. Chem. 281: 4173-4182 [Abstract] [Full Text]  
  • Lu, R., Ghory, H. Z., Engelman, A. (2005). Genetic Analyses of Conserved Residues in the Carboxyl-Terminal Domain of Human Immunodeficiency Virus Type 1 Integrase. J. Virol. 79: 10356-10368 [Abstract] [Full Text]  
  • Busschots, K., Vercammen, J., Emiliani, S., Benarous, R., Engelborghs, Y., Christ, F., Debyser, Z. (2005). The Interaction of LEDGF/p75 with Integrase Is Lentivirus-specific and Promotes DNA Binding. J. Biol. Chem. 280: 17841-17847 [Abstract] [Full Text]  
  • Hehl, E. A., Joshi, P., Kalpana, G. V., Prasad, V. R. (2004). Interaction between Human Immunodeficiency Virus Type 1 Reverse Transcriptase and Integrase Proteins. J. Virol. 78: 5056-5067 [Abstract] [Full Text]  
  • Svarovskaia, E. S., Barr, R., Zhang, X., Pais, G. C. G., Marchand, C., Pommier, Y., Burke, T. R. Jr., Pathak, V. K. (2004). Azido-Containing Diketo Acid Derivatives Inhibit Human Immunodeficiency Virus Type 1 Integrase In Vivo and Influence the Frequency of Deletions at Two-Long-Terminal-Repeat-Circle Junctions. J. Virol. 78: 3210-3222 [Abstract] [Full Text]  
  • Calmels, C., de Soultrait, V. R., Caumont, A., Desjobert, C., Faure, A., Fournier, M., Tarrago-Litvak, L., Parissi, V. (2004). Biochemical and random mutagenesis analysis of the region carrying the catalytic E152 amino acid of HIV-1 integrase. Nucleic Acids Res 32: 1527-1538 [Abstract] [Full Text]  
  • Devroe, E., Engelman, A., Silver, P. A. (2003). Intracellular transport of human immunodeficiency virus type 1 integrase. J. Cell Sci. 116: 4401-4408 [Abstract] [Full Text]  
  • Hartlieb, B., Modrof, J., Muhlberger, E., Klenk, H.-D., Becker, S. (2003). Oligomerization of Ebola Virus VP30 Is Essential for Viral Transcription and Can Be Inhibited by a Synthetic Peptide. J. Biol. Chem. 278: 41830-41836 [Abstract] [Full Text]  
  • Chiu, R., Grandgenett, D. P. (2003). Molecular and Genetic Determinants of Rous Sarcoma Virus Integrase for Concerted DNA Integration. J. Virol. 77: 6482-6492 [Abstract] [Full Text]  
  • Yi, J., Cheng, H., Andrake, M. D., Dunbrack, R. L. Jr., Roder, H., Skalka, A. M. (2002). Mapping the Epitope of an Inhibitory Monoclonal Antibody to the C-terminal DNA-binding Domain of HIV-1 Integrase. J. Biol. Chem. 277: 12164-12174 [Abstract] [Full Text]  
  • Appa, R. S., Shin, C.-G., Lee, P., Chow, S. A. (2001). 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. 276: 45848-45855 [Abstract] [Full Text]  
  • Chiu, R., Grandgenett, D. P. (2000). Avian Retrovirus DNA Internal Attachment Site Requirements for Full-Site Integration In Vitro. J. Virol. 74: 8292-8298 [Abstract] [Full Text]  
  • Chen, J. C.-H., Krucinski, J., Miercke, L. J. W., Finer-Moore, J. S., Tang, A. H., Leavitt, A. D., Stroud, R. M. (2000). Crystal structure of the HIV-1 integrase catalytic core and C-terminal domains: A model for viral DNA binding. Proc. Natl. Acad. Sci. USA 10.1073/pnas.150220297v1 [Abstract] [Full Text]  
  • Hindmarsh, P., Leis, J. (1999). Retroviral DNA Integration. Microbiol. Mol. Biol. Rev. 63: 836-843 [Abstract] [Full Text]  
  • Ishikawa, T., Okui, N., Kobayashi, N., Sakuma, R., Kitamura, T., Kitamura, Y. (1999). Monoclonal Antibodies against the Minimal DNA-Binding Domain in the Carboxyl-Terminal Region of Human Immunodeficiency Virus Type 1 Integrase. J. Virol. 73: 4475-4480 [Abstract] [Full Text]  
  • van den Ent, F. M. I., Vos, A., Plasterk, R. H. A. (1999). Dissecting the Role of the N-Terminal Domain of Human Immunodeficiency Virus Integrase by trans-Complementation Analysis. J. Virol. 73: 3176-3183 [Abstract] [Full Text]  
  • Uhrig, J. F., Soellick, T.-R., Minke, C. J., Philipp, C., Kellmann, J.-W., Schreier, P. H. (1999). Homotypic interaction and multimerization of nucleocapsid protein of tomato spotted wilt tospovirus: Identification and characterization of two interacting domains. Proc. Natl. Acad. Sci. USA 96: 55-60 [Abstract] [Full Text]  
  • Chen, J. C.-H., Krucinski, J., Miercke, L. J. W., Finer-Moore, J. S., Tang, A. H., Leavitt, A. D., Stroud, R. M. (2000). Crystal structure of the HIV-1 integrase catalytic core and C-terminal domains: A model for viral DNA binding. Proc. Natl. Acad. Sci. USA 97: 8233-8238 [Abstract] [Full Text]  
  • Deprez, E., Tauc, P., Leh, H., Mouscadet, J.-F., Auclair, C., Hawkins, M. E., Brochon, J.-C. (2001). DNA binding induces dissociation of the multimeric form of HIV-1 integrase: A time-resolved fluorescence anisotropy study. Proc. Natl. Acad. Sci. USA 98: 10090-10095 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»