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 Welker, R.
Right arrow Articles by Kräusslich, H.-G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Welker, R.
Right arrow Articles by Kräusslich, H.-G.

 Previous Article  |  Next Article 

Journal of Virology, February 2000, p. 1168-1177, Vol. 74, No. 3
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Biochemical and Structural Analysis of Isolated Mature Cores of Human Immunodeficiency Virus Type 1

Reinhold Welker,dagger Heinrich Hohenberg, Uwe Tessmer, Carola Huckhagel, and Hans-Georg Kräusslich*

Heinrich-Pette-Institut für experimentelle Virologie und Immunologie an der Universität Hamburg, D-20251 Hamburg, Germany

Received 3 August 1999/Accepted 4 November 1999

Mature human immunodeficiency virus type 1 (HIV-1) particles contain a cone-shaped core structure consisting of the internal ribonucleoprotein complex encased in a proteinaceous shell derived from the viral capsid protein. Because of their very low stability after membrane removal, HIV-1 cores have not been purified in quantities sufficient for structural and biochemical analysis. Based on our in vitro assembly experiments, we have developed a novel method for isolation of intact mature HIV-1 cores. Concentrated virus suspensions were briefly treated with nonionic detergent and immediately centrifuged in a microcentrifuge for short periods of time. The resuspended pellet was subsequently analyzed by negative-stain and thin-section electron microscopy and by immunoelectron microscopy. Abundant cone-shaped cores as well as tubular and aberrant structures were observed. Stereo images showed that core structures preserved their three-dimensional architecture and exhibited a regular substructure. Detailed analysis of 155 cores revealed an average length of ca. 103 nm, an average diameter at the base of ca. 52 nm, and an average angle of 21.3°. There was significant variability in all parameters, indicating that HIV cores are not homogeneous. Immunoblot analysis of core preparations allowed semiquantitative estimation of the relative amounts of viral and cellular proteins inside the HIV-1 core, yielding a model for the topology of various proteins inside the virion.

* Corresponding author. Mailing address: Heinrich-Pette-Institut für experimentelle Virologie und Immunologie an der Universität Hamburg, Martinistr. 52, D-20251 Hamburg, Germany. Phone: 49 40 48051-241. Fax: 49 40 48051-184. E-mail: hgk{at}hpi.uni-hamburg.de.

dagger Present address: Department of Molecular Genetics and Microbiology, State University of New York, Stony Brook, NY 11794-5222.

Journal of Virology, February 2000, p. 1168-1177, Vol. 74, No. 3
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Sebastian, S., Grutter, C., de Castillia, C. S., Pertel, T., Olivari, S., Grutter, M. G., Luban, J. (2009). An Invariant Surface Patch on the TRIM5{alpha} PRYSPRY Domain Is Required for Retroviral Restriction but Dispensable for Capsid Binding. J. Virol. 83: 3365-3373 [Abstract] [Full Text]  
  • Laguette, N., Benichou, S., Basmaciogullari, S. (2009). Human Immunodeficiency Virus Type 1 Nef Incorporation into Virions Does Not Increase Infectivity. J. Virol. 83: 1093-1104 [Abstract] [Full Text]  
  • Langelier, C. R., Sandrin, V., Eckert, D. M., Christensen, D. E., Chandrasekaran, V., Alam, S. L., Aiken, C., Olsen, J. C., Kar, A. K., Sodroski, J. G., Sundquist, W. I. (2008). Biochemical Characterization of a Recombinant TRIM5{alpha} Protein That Restricts Human Immunodeficiency Virus Type 1 Replication. J. Virol. 82: 11682-11694 [Abstract] [Full Text]  
  • Purdy, J. G., Flanagan, J. M., Ropson, I. J., Rennoll-Bankert, K. E., Craven, R. C. (2008). Critical Role of Conserved Hydrophobic Residues within the Major Homology Region in Mature Retroviral Capsid Assembly. J. Virol. 82: 5951-5961 [Abstract] [Full Text]  
  • Coren, L. V., Thomas, J. A., Chertova, E., Sowder, R. C. II, Gagliardi, T. D., Gorelick, R. J., Ott, D. E. (2007). Mutational Analysis of the C-Terminal Gag Cleavage Sites in Human Immunodeficiency Virus Type 1. J. Virol. 81: 10047-10054 [Abstract] [Full Text]  
  • Haim, H., Steiner, I., Panet, A. (2007). Time Frames for Neutralization during the Human Immunodeficiency Virus Type 1 Entry Phase, as Monitored in Synchronously Infected Cell Cultures. J. Virol. 81: 3525-3534 [Abstract] [Full Text]  
  • Darnell, G. A., Schroder, W. A., Gardner, J., Harrich, D., Yu, H., Medcalf, R. L., Warrilow, D., Antalis, T. M., Sonza, S., Suhrbier, A. (2006). SerpinB2 Is an Inducible Host Factor Involved in Enhancing HIV-1 Transcription and Replication. J. Biol. Chem. 281: 31348-31358 [Abstract] [Full Text]  
  • Roy, B. B., Hu, J., Guo, X., Russell, R. S., Guo, F., Kleiman, L., Liang, C. (2006). Association of RNA Helicase A with Human Immunodeficiency Virus Type 1 Particles. J. Biol. Chem. 281: 12625-12635 [Abstract] [Full Text]  
  • Stremlau, M., Perron, M., Lee, M., Li, Y., Song, B., Javanbakht, H., Diaz-Griffero, F., Anderson, D. J., Sundquist, W. I., Sodroski, J. (2006). From the Cover: Specific recognition and accelerated uncoating of retroviral capsids by the TRIM5{alpha} restriction factor. Proc. Natl. Acad. Sci. USA 103: 5514-5519 [Abstract] [Full Text]  
  • Davis, M. R., Jiang, J., Zhou, J., Freed, E. O., Aiken, C. (2006). A Mutation in the Human Immunodeficiency Virus Type 1 Gag Protein Destabilizes the Interaction of the Envelope Protein Subunits gp120 and gp41. J. Virol. 80: 2405-2417 [Abstract] [Full Text]  
  • Brugger, B., Glass, B., Haberkant, P., Leibrecht, I., Wieland, F. T., Krausslich, H.-G. (2006). The HIV lipidome: A raft with an unusual composition. Proc. Natl. Acad. Sci. USA 103: 2641-2646 [Abstract] [Full Text]  
  • Galetto, R., Giacomoni, V., Veron, M., Negroni, M. (2006). Dissection of a Circumscribed Recombination Hot Spot in HIV-1 after a Single Infectious Cycle. J. Biol. Chem. 281: 2711-2720 [Abstract] [Full Text]  
  • Fossen, T., Wray, V., Bruns, K., Rachmat, J., Henklein, P., Tessmer, U., Maczurek, A., Klinger, P., Schubert, U. (2005). Solution Structure of the Human Immunodeficiency Virus Type 1 p6 Protein. J. Biol. Chem. 280: 42515-42527 [Abstract] [Full Text]  
  • Gurer, C., Hoglund, A., Hoglund, S., Luban, J. (2005). ATP{gamma}S Disrupts Human Immunodeficiency Virus Type 1 Virion Core Integrity. J. Virol. 79: 5557-5567 [Abstract] [Full Text]  
  • Scholz, I., Arvidson, B., Huseby, D., Barklis, E. (2005). Virus Particle Core Defects Caused by Mutations in the Human Immunodeficiency Virus Capsid N-Terminal Domain. J. Virol. 79: 1470-1479 [Abstract] [Full Text]  
  • Daecke, J., Fackler, O. T., Dittmar, M. T., Krausslich, H.-G. (2005). Involvement of Clathrin-Mediated Endocytosis in Human Immunodeficiency Virus Type 1 Entry. J. Virol. 79: 1581-1594 [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]  
  • Stuchell, M. D., Garrus, J. E., Muller, B., Stray, K. M., Ghaffarian, S., McKinnon, R., Krausslich, H.-G., Morham, S. G., Sundquist, W. I. (2004). The Human Endosomal Sorting Complex Required for Transport (ESCRT-I) and Its Role in HIV-1 Budding. J. Biol. Chem. 279: 36059-36071 [Abstract] [Full Text]  
  • Ganser-Pornillos, B. K., von Schwedler, U. K., Stray, K. M., Aiken, C., Sundquist, W. I. (2004). Assembly Properties of the Human Immunodeficiency Virus Type 1 CA Protein. J. Virol. 78: 2545-2552 [Abstract] [Full Text]  
  • Guo, X., Hu, J., Whitney, J. B., Russell, R. S., Liang, C. (2004). Important Role for the CA-NC Spacer Region in the Assembly of Bovine Immunodeficiency Virus Gag Protein. J. Virol. 78: 551-560 [Abstract] [Full Text]  
  • Tang, S., Murakami, T., Cheng, N., Steven, A. C., Freed, E. O., Levin, J. G. (2003). Human Immunodeficiency Virus Type 1 N-Terminal Capsid Mutants Containing Cores with Abnormally High Levels of Capsid Protein and Virtually No Reverse Transcriptase. J. Virol. 77: 12592-12602 [Abstract] [Full Text]  
  • Tobiume, M., Lineberger, J. E., Lundquist, C. A., Miller, M. D., Aiken, C. (2003). Nef Does Not Affect the Efficiency of Human Immunodeficiency Virus Type 1 Fusion with Target Cells. J. Virol. 77: 10645-10650 [Abstract] [Full Text]  
  • Ott, D. E., Coren, L. V., Chertova, E. N., Gagliardi, T. D., Nagashima, K., Sowder, R. C. II, Poon, D. T. K., Gorelick, R. J. (2003). Elimination of Protease Activity Restores Efficient Virion Production to a Human Immunodeficiency Virus Type 1 Nucleocapsid Deletion Mutant. J. Virol. 77: 5547-5556 [Abstract] [Full Text]  
  • von Schwedler, U. K., Stray, K. M., Garrus, J. E., Sundquist, W. I. (2003). Functional Surfaces of the Human Immunodeficiency Virus Type 1 Capsid Protein. J. Virol. 77: 5439-5450 [Abstract] [Full Text]  
  • Moumen, A., Polomack, L., Unge, T., Veron, M., Buc, H., Negroni, M. (2003). Evidence for a Mechanism of Recombination during Reverse Transcription Dependent on the Structure of the Acceptor RNA. J. Biol. Chem. 278: 15973-15982 [Abstract] [Full Text]  
  • Forshey, B. M., Aiken, C. (2003). Disassembly of Human Immunodeficiency Virus Type 1 Cores In Vitro Reveals Association of Nef with the Subviral Ribonucleoprotein Complex. J. Virol. 77: 4409-4414 [Abstract] [Full Text]  
  • Hoglund, S., Su, J., Reneby, S. S., Vegvari, A., Hjerten, S., Sintorn, I.-M., Foster, H., Wu, Y.-P., Nystrom, I., Vahlne, A. (2002). Tripeptide Interference with Human Immunodeficiency Virus Type 1 Morphogenesis. Antimicrob. Agents Chemother. 46: 3597-3605 [Abstract] [Full Text]  
  • Johnson, M. C., Scobie, H. M., Ma, Y. M., Vogt, V. M. (2002). Nucleic Acid-Independent Retrovirus Assembly Can Be Driven by Dimerization. J. Virol. 76: 11177-11185 [Abstract] [Full Text]  
  • Forshey, B. M., von Schwedler, U., Sundquist, W. I., Aiken, C. (2002). Formation of a Human Immunodeficiency Virus Type 1 Core of Optimal Stability Is Crucial for Viral Replication. J. Virol. 76: 5667-5677 [Abstract] [Full Text]  
  • Muller, B., Patschinsky, T., Krausslich, H.-G. (2002). The Late-Domain-Containing Protein p6 Is the Predominant Phosphoprotein of Human Immunodeficiency Virus Type 1 Particles. J. Virol. 76: 1015-1024 [Abstract] [Full Text]  
  • Jenkins, Y., Pornillos, O., Rich, R. L., Myszka, D. G., Sundquist, W. I., Malim, M. H. (2001). Biochemical Analyses of the Interactions between Human Immunodeficiency Virus Type 1 Vpr and p6Gag. J. Virol. 75: 10537-10542 [Abstract] [Full Text]  
  • Tang, S., Murakami, T., Agresta, B. E., Campbell, S., Freed, E. O., Levin, J. G. (2001). Human Immunodeficiency Virus Type 1 N-Terminal Capsid Mutants That Exhibit Aberrant Core Morphology and Are Blocked in Initiation of Reverse Transcription in Infected Cells. J. Virol. 75: 9357-9366 [Abstract] [Full Text]  
  • Yovandich, J. L., Chertova, E. N., Kane, B. P., Gagliardi, T. D., Bess, J. W. Jr., Sowder, R. C. II, Henderson, L. E., Gorelick, R. J. (2001). Alteration of Zinc-Binding Residues of Simian Immunodeficiency Virus p8NC Results in Subtle Differences in Gag Processing and Virion Maturation Associated with Degradative Loss of Mutant NC. J. Virol. 75: 115-124 [Abstract] [Full Text]  
  • Öhagen, A., Gabuzda, D. (2000). Role of Vif in Stability of the Human Immunodeficiency Virus Type 1 Core. J. Virol. 74: 11055-11066 [Abstract] [Full Text]  
  • Wyma, D. J., Kotov, A., Aiken, C. (2000). Evidence for a Stable Interaction of gp41 with Pr55Gag in Immature Human Immunodeficiency Virus Type 1 Particles. J. Virol. 74: 9381-9387 [Abstract] [Full Text]  
  • Müller, B., Tessmer, U., Schubert, U., Kräusslich, H.-G. (2000). Human Immunodeficiency Virus Type 1 Vpr Protein Is Incorporated into the Virion in Significantly Smaller Amounts than Gag and Is Phosphorylated in Infected Cells. J. Virol. 74: 9727-9731 [Abstract] [Full Text]  
  • Accola, M. A., Öhagen, A., Göttlinger, H. G. (2000). Isolation of Human Immunodeficiency Virus Type 1 Cores: Retention of Vpr in the Absence of p6gag. J. Virol. 74: 6198-6202 [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»