Double-labelled HIV-1 particles for study of virus-cell interaction.
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.
Lampe
Marko
M
Department of Virology, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany.
Briggs
John A G
JA
Endress
Thomas
T
Glass
Bärbel
B
Riegelsberger
Stefan
S
Kräusslich
Hans-Georg
HG
Lamb
Don C
DC
Bräuchle
Christoph
C
Müller
Barbara
B
D001692:Biological Transport; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D016336:Gene Products, vpr / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D013194:Staining and Labeling; D014763:Viral Matrix Proteins / Q000378:metabolism; D014773:Virology / Q000379:methods; D014779:Virus Replication; D054325:vpr Gene Products, Human Immunodeficiency Virus; D000097573:Red Fluorescent Protein
D016336:Gene Products, vpr; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D014763:Viral Matrix Proteins; D054325:vpr Gene Products, Human Immunodeficiency Virus; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein