J. Virol., 07 1996, 4299-4310, Vol 70, No. 7
Copyright © 1996, American Society for Microbiology

Binding of the human immunodeficiency virus type 1 Gag polyprotein to cyclophilin A is mediated by the central region of capsid and requires Gag dimerization

J Colgan, HE Yuan, EK Franke and J Luban
Department of Microbiology, College of Physicians and Surgeons of Columbia University, New York, New York 10032USA.

The cellular peptidyl-prolyl isomerase cyclophilin A (CyPA) is incorporated into human immunodeficiency virus type 1 (HIV-1) virions via direct contacts with the HIV-1 Gag polyprotein. Disruption of the Gag-CyPA interaction leads to the production of HIV-1 particles lacking CyPA; these virions are noninfectious, indicating that contacts between CyPA and Gag are necessary for HIV-1 replication. Here, we have used the yeast two-hybrid system in conjunction with an in vitro binding assay to identify the minimal domain of Gag required for binding to CyPA. Analysis of a panel of gag deletion mutants in the two-hybrid system indicated that a region spanning the central portion of the capsid (CA) domain was sufficient for interactions with CyPA, but discrepancies between results obtained in different fusion protein contexts suggested that multimerization of Gag might also be necessary for binding to CyPA. Consistent with a requirement for multimerization, the binding of Gag to CyPA in vitro required a region within the nucleocapsid (NC) domain shown previously to be important for Gag self- association. Substitution of a heterologous dimerization motif for the region from NC also promoted specific binding to CyPA, confirming that interactions with CyPA are dependent on Gag multimerization. Fusion of the heterologous dimerization motif to a 100-amino-acid domain from CA was sufficient for binding to CyPA in vitro. These results define the minimal CyPA-binding domain within Gag and provide insight into the mechanism by which CyPA is incorporated into HIV-1 virions.

This article has been cited by other articles:

• Luban, J. (2007). Cyclophilin A, TRIM5, and Resistance to Human Immunodeficiency Virus Type 1 Infection. J. Virol. 81: 1054-1061 [Full Text]  
• Ribeiro, I. P., Menezes, A. N., Moreira, M. A. M., Bonvicino, C. R., Seuanez, H. N., Soares, M. A. (2005). Evolution of Cyclophilin A and TRIMCyp Retrotransposition in New World Primates. J. Virol. 79: 14998-15003 [Abstract] [Full Text]  
• Berthoux, L., Sebastian, S., Sayah, D. M., Luban, J. (2005). Disruption of Human TRIM5{alpha} Antiviral Activity by Nonhuman Primate Orthologues. J. Virol. 79: 7883-7888 [Abstract] [Full Text]  
• Khan, M., Jin, L., Miles, L., Bond, V. C., Powell, M. D. (2005). Chimeric Human Immunodeficiency Virus Type 1 Virions That Contain the Simian Immunodeficiency Virus nef Gene Are Cyclosporin A Resistant. J. Virol. 79: 3211-3216 [Abstract] [Full Text]  
• Bon Homme, M., Carter, C., Scarlata, S. (2005). The Cysteine Residues of HIV-1 Capsid Regulate Oligomerization and Cyclophilin A-Induced Changes. Biophys. J 88: 2078-2088 [Abstract] [Full Text]  
• Gurer, C., Berthoux, L., Luban, J. (2005). Covalent Modification of Human Immunodeficiency Virus Type 1 p6 by SUMO-1. J. Virol. 79: 910-917 [Abstract] [Full Text]  
• Sokolskaja, E., Sayah, D. M., Luban, J. (2004). Target Cell Cyclophilin A Modulates Human Immunodeficiency Virus Type 1 Infectivity. J. Virol. 78: 12800-12808 [Abstract] [Full Text]  
• Khan, M., Jin, L., Huang, M. B., Miles, L., Bond, V. C., Powell, M. D. (2004). Chimeric Human Immunodeficiency Virus Type 1 (HIV-1) Virions Containing HIV-2 or Simian Immunodeficiency Virus Nef Are Resistant to Cyclosporine Treatment. J. Virol. 78: 1843-1850 [Abstract] [Full Text]  
• Castro, A. P. V., Carvalho, T. M. U., Moussatche, N., Damaso, C. R. A. (2003). Redistribution of Cyclophilin A to Viral Factories during Vaccinia Virus Infection and Its Incorporation into Mature Particles. J. Virol. 77: 9052-9068 [Abstract] [Full Text]  
• Bose, S., Mathur, M., Bates, P., Joshi, N., Banerjee, A. K. (2003). Requirement for cyclophilin A for the replication of vesicular stomatitis virus New Jersey serotype. J. Gen. Virol. 84: 1687-1699 [Abstract] [Full Text]  
• Khan, M., Garcia-Barrio, M., Powell, M. D. (2003). Treatment of Human Immunodeficiency Virus Type 1 Virions Depleted of Cyclophilin A by Natural Endogenous Reverse Transcription Restores Infectivity. J. Virol. 77: 4431-4434 [Abstract] [Full Text]  
• Yoshimatsu, K., Lee, B.-H., Araki, K., Morimatsu, M., Ogino, M., Ebihara, H., Arikawa, J. (2002). The Multimerization of Hantavirus Nucleocapsid Protein Depends on Type-Specific Epitopes. J. Virol. 77: 943-952 [Abstract] [Full Text]  
• Saphire, A. C. S., Bobardt, M. D., Gallay, P. A. (2002). Cyclophilin A Plays Distinct Roles in Human Immunodeficiency Virus Type 1 Entry and Postentry Events, as Revealed by Spinoculation. J. Virol. 76: 4671-4677 [Abstract] [Full Text]  
• Saphire, A. C. S., Bobardt, M. D., Gallay, P. A. (2002). trans-Complementation Rescue of Cyclophilin A-Deficient Viruses Reveals that the Requirement for Cyclophilin A in Human Immunodeficiency Virus Type 1 Replication Is Independent of Its Isomerase Activity. J. Virol. 76: 2255-2262 [Abstract] [Full Text]  
• Cimarelli, A., Sandin, S., Höglund, S., Luban, J. (2000). Basic Residues in Human Immunodeficiency Virus Type 1 Nucleocapsid Promote Virion Assembly via Interaction with RNA. J. Virol. 74: 3046-3057 [Abstract] [Full Text]  
• Burniston, M. T., Cimarelli, A., Colgan, J., Curtis, S. P., Luban, J. (1999). Human Immunodeficiency Virus Type 1 Gag Polyprotein Multimerization Requires the Nucleocapsid Domain and RNA and Is Promoted by the Capsid-Dimer Interface and the Basic Region of Matrix Protein. J. Virol. 73: 8527-8540 [Abstract] [Full Text]  
• Cimarelli, A., Luban, J. (1999). Translation Elongation Factor 1-Alpha Interacts Specifically with the Human Immunodeficiency Virus Type 1 Gag Polyprotein. J. Virol. 73: 5388-5401 [Abstract] [Full Text]  
• Srivastava, M., Cartas, M., Rizvi, T. A., Singh, S. P., Serio, D., Kalyanaraman, V. S., Pollard, H. B., Srinivasan, A. (1999). HIV-1 Gag shares a signature motif with annexin (Anx7), which is required for virus replication. Proc. Natl. Acad. Sci. USA 96: 2704-2709 [Abstract] [Full Text]  
• Sharma, U. K., Trujillo, J., Song, H.-F., Saitta, F. P., Laeyendecker, O. B., Castillo, R., Arango-Jaramillo, S., Sridharan, G., Dettenhofer, M., Blakemore, K., Yu, X.-F., Schwartz, D. H. (1998). A Novel Factor Produced by Placental Cells with Activity Against HIV-1. J. Immunol. 161: 6406-6412 [Abstract] [Full Text]  
• de Rocquigny, H., Petitjean, P., Tanchou, V., Decimo, D., Drouot, L., Delaunay, T., Darlix, J.-L., Roques, B. P. (1997). The Zinc Fingers of HIV Nucleocapsid Protein NCp7 Direct Interactions with the Viral Regulatory Protein Vpr. J. Biol. Chem. 272: 30753-30759 [Abstract] [Full Text]  
• Cohen, G. B., Rangan, V. S., Chen, B. K., Smith, S., Baltimore, D. (2000). The Human Thioesterase II Protein Binds to a Site on HIV-1 Nef Critical for CD4 Down-regulation. J. Biol. Chem. 275: 23097-23105 [Abstract] [Full Text]  
• Pushkarsky, T., Zybarth, G., Dubrovsky, L., Yurchenko, V., Tang, H., Guo, H., Toole, B., Sherry, B., Bukrinsky, M. (2001). CD147 facilitates HIV-1 infection by interacting with virus-associated cyclophilin A. Proc. Natl. Acad. Sci. USA 98: 6360-6365 [Abstract] [Full Text]