Document Detail

Double-labeled rabies virus: live tracking of enveloped virus transport.
MedLine Citation:
PMID:  17928343     Owner:  NLM     Status:  MEDLINE    
Here we describe a strategy to fluorescently label the envelope of rabies virus (RV), of the Rhabdoviridae family, in order to track the transport of single enveloped viruses in living cells. Red fluorescent proteins (tm-RFP) were engineered to comprise the N-terminal signal sequence and C-terminal transmembrane spanning and cytoplasmic domain sequences of the RV glycoprotein (G). Two variants of tm-RFP were transported to and anchored in the cell surface membrane, independent of glycosylation. As shown by confocal microscopy, tm-RFP colocalized at the cell surface with the RV matrix and G protein and was incorporated into G gene-deficient virus particles. Recombinant RV expressing the membrane-anchored tm-RFP in addition to G yielded infectious viruses with mosaic envelopes containing both tm-RFP and G. Viable double-labeled virus particles comprising a red fluorescent envelope and a green fluorescent ribonucleoprotein were generated by expressing in addition an enhanced green fluorescent protein-phosphoprotein fusion construct (S. Finke, K. Brzozka, and K. K. Conzelmann, J. Virol. 78:12333-12343, 2004). Individual enveloped virus particles were observed under live cell conditions as extracellular particles and inside endosomal vesicles. Importantly, double-labeled RVs were transported in the retrograde direction over long distances in neurites of in vitro-differentiated NS20Y neuroblastoma cells. This indicates that the typical retrograde axonal transport of RV to the central nervous system involves neuronal transport vesicles in which complete enveloped RV particles are carried as a cargo.
Yvonne Klingen; Karl-Klaus Conzelmann; Stefan Finke
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-10-10
Journal Detail:
Title:  Journal of virology     Volume:  82     ISSN:  1098-5514     ISO Abbreviation:  J. Virol.     Publication Date:  2008 Jan 
Date Detail:
Created Date:  2007-12-11     Completed Date:  2008-01-10     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  237-45     Citation Subset:  IM    
Friedrich-Loeffler-Institute, Boddenblick 5a, D-17493 Greifswald-Riems, Germany.
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MeSH Terms
Antigens, Viral / genetics,  metabolism
Cell Line
Cell Membrane / chemistry
Endosomes / virology
Genes, Reporter*
Glycoproteins / genetics,  metabolism
Green Fluorescent Proteins / genetics,  metabolism
Luminescent Proteins / genetics,  metabolism
Microscopy, Confocal
Neurons / virology
Phosphoproteins / genetics,  metabolism
Rabies virus / genetics,  physiology*
Recombinant Fusion Proteins / genetics,  metabolism
Ribonucleoproteins / genetics,  metabolism
Staining and Labeling / methods*
Viral Envelope Proteins / genetics,  metabolism
Viral Matrix Proteins / metabolism
Viral Structural Proteins / genetics,  metabolism
Virion / chemistry
Virus Assembly / physiology
Reg. No./Substance:
0/Antigens, Viral; 0/Glycoproteins; 0/Luminescent Proteins; 0/P phosphoprotein, Rabies virus; 0/Phosphoproteins; 0/Recombinant Fusion Proteins; 0/Ribonucleoproteins; 0/Viral Envelope Proteins; 0/Viral Matrix Proteins; 0/Viral Structural Proteins; 0/enhanced green fluorescent protein; 0/glycoprotein G, Rabies virus; 0/red fluorescent protein; 147336-22-9/Green Fluorescent Proteins

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