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Title: Double-labelled HIV-1 particles for study of virus-cell interaction

Abstract

Human immunodeficiency virus (HIV) delivers its genome to a host cell through fusion of the viral envelope with a cellular membrane. While the viral and cellular proteins involved in entry have been analyzed in detail, the dynamics of virus-cell fusion are largely unknown. Single virus tracing (SVT) provides the unique opportunity to visualize viral particles in real time allowing direct observation of the dynamics of this stochastic process. For this purpose, we developed a double-coloured HIV derivative carrying a green fluorescent label attached to the viral matrix protein combined with a red label fused to the viral Vpr protein designed to distinguish between complete virions and subviral particles lacking MA after membrane fusion. We present here a detailed characterization of this novel tool together with exemplary live cell imaging studies, demonstrating its suitability for real-time analyses of HIV-cell interaction.

Authors:
 [1];  [2];  [2];  [1];  [2];  [1];  [3];  [4];  [5];  [3];  [5];  [6]
  1. Department of Virology, Universitaetsklinikum Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg (Germany)
  2. Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich (Germany)
  3. Center for NanoScience, Ludwig-Maximilians-Universitaet Muenchen, Geschwister-Scholl-Platz 1, 80539 Munich (Germany)
  4. (United States)
  5. (Germany)
  6. Department of Virology, Universitaetsklinikum Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg (Germany). E-mail: Barbara_Mueller@med.uni-heidelberg.de
Publication Date:
OSTI Identifier:
20977002
Resource Type:
Journal Article
Resource Relation:
Journal Name: Virology; Journal Volume: 360; Journal Issue: 1; Other Information: DOI: 10.1016/j.virol.2006.10.005; PII: S0042-6822(06)00738-0; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AIDS VIRUS; CELL MEMBRANES; FLUORESCENCE; LABELLING; PROTEINS; STOCHASTIC PROCESSES; SULFATES

Citation Formats

Lampe, Marko, Briggs, John A.G., Endress, Thomas, Glass, Baerbel, Riegelsberger, Stefan, Kraeusslich, Hans-Georg, Lamb, Don C., Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich, Braeuchle, Christoph, Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich, and Mueller, Barbara. Double-labelled HIV-1 particles for study of virus-cell interaction. United States: N. p., 2007. Web. doi:10.1016/j.virol.2006.10.005.
Lampe, Marko, Briggs, John A.G., Endress, Thomas, Glass, Baerbel, Riegelsberger, Stefan, Kraeusslich, Hans-Georg, Lamb, Don C., Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich, Braeuchle, Christoph, Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich, & Mueller, Barbara. Double-labelled HIV-1 particles for study of virus-cell interaction. United States. doi:10.1016/j.virol.2006.10.005.
Lampe, Marko, Briggs, John A.G., Endress, Thomas, Glass, Baerbel, Riegelsberger, Stefan, Kraeusslich, Hans-Georg, Lamb, Don C., Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich, Braeuchle, Christoph, Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich, and Mueller, Barbara. Fri . "Double-labelled HIV-1 particles for study of virus-cell interaction". United States. doi:10.1016/j.virol.2006.10.005.
@article{osti_20977002,
title = {Double-labelled HIV-1 particles for study of virus-cell interaction},
author = {Lampe, Marko and Briggs, John A.G. and Endress, Thomas and Glass, Baerbel and Riegelsberger, Stefan and Kraeusslich, Hans-Georg and Lamb, Don C. and Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801 and Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich and Braeuchle, Christoph and Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universitaet Muenchen, Butenandtstr. 11, Haus E, 81377 Munich and Mueller, Barbara},
abstractNote = {Human immunodeficiency virus (HIV) delivers its genome to a host cell through fusion of the viral envelope with a cellular membrane. While the viral and cellular proteins involved in entry have been analyzed in detail, the dynamics of virus-cell fusion are largely unknown. Single virus tracing (SVT) provides the unique opportunity to visualize viral particles in real time allowing direct observation of the dynamics of this stochastic process. For this purpose, we developed a double-coloured HIV derivative carrying a green fluorescent label attached to the viral matrix protein combined with a red label fused to the viral Vpr protein designed to distinguish between complete virions and subviral particles lacking MA after membrane fusion. We present here a detailed characterization of this novel tool together with exemplary live cell imaging studies, demonstrating its suitability for real-time analyses of HIV-cell interaction.},
doi = {10.1016/j.virol.2006.10.005},
journal = {Virology},
number = 1,
volume = 360,
place = {United States},
year = {Fri Mar 30 00:00:00 EDT 2007},
month = {Fri Mar 30 00:00:00 EDT 2007}
}
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