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Published online 4 November 2002. doi:10.1083/jcb.200203150
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© The Rockefeller University Press, 0021-9525/2002/11/441 $5.00
The Journal of Cell Biology, Volume 159, Number 3, 441-452

Article

 Visualization of the intracellular behavior of HIV in living cells

David McDonald1, Marie A. Vodicka2, Ginger Lucero3, Tatyana M. Svitkina4, Gary G. Borisy4, Michael Emerman2 and Thomas J. Hope1

1 University of Illinois, Chicago, IL 60612
2 Fred Hutchinson Cancer Research Center, Seattle, WA 98109
3 The Salk Institute, La Jolla, CA 92186
4 Northwestern University Medical School, Chicago, IL 60611

Address correspondence to Thomas J. Hope, Dept. of Microbiology and Immunology, MSB E-704, M/C 790, 835 South Wolcott Ave., Chicago, IL 60612. Tel.: (312) 413-3424. Fax: (312) 996-6415. E-mail: thope{at}uic.edu

To track the behavior of human immunodeficiency virus (HIV)-1 in the cytoplasm of infected cells, we have tagged virions by incorporation of HIV Vpr fused to the GFP. Observation of the GFP-labeled particles in living cells revealed that they moved in curvilinear paths in the cytoplasm and accumulated in the perinuclear region, often near the microtubule-organizing center. Further studies show that HIV uses cytoplasmic dynein and the microtubule network to migrate toward the nucleus. By combining GFP fused to the NH2 terminus of HIV-1 Vpr tagging with other labeling techniques, it was possible to determine the state of progression of individual particles through the viral life cycle. Correlation of immunofluorescent and electron micrographs allowed high resolution imaging of microtubule-associated structures that are proposed to be reverse transcription complexes. Based on these observations, we propose that HIV uses dynein and the microtubule network to facilitate the delivery of the viral genome to the nucleus of the cell during early postentry steps of the HIV life cycle.

Key Words: HIV-1; reverse transcription complex; dynein; fluorescent microscopy; electron microscopy


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