© The Rockefeller University Press,
0021-9525/1998//875 $5.00
The Journal of Cell Biology, Volume 143, Number 4,
, 1998 875-885
Characterization of HIV-1 Vpr Nuclear Import: Analysis of Signals and Pathways
Yonchu Jenkins*,
Michele McEntee*,
Karsten Weis
, and
Warner C. Greene*,
,||
* Gladstone Institute of Virology and Immunology,
Department of Biochemistry and Biophysics,
Department of Medicine, and || Department of Microbiology and Immunology, University of California, San Francisco, California 94141-9100
While the Vpr protein of HIV-1 has been implicated in import of the viral preintegration complex across the nuclear pore complex (NPC) of nondividing cellular hosts, the mechanism by which Vpr enters the nucleus remains unknown. We now demonstrate that Vpr contains two discrete nuclear targeting signals that use two different import pathways, both of which are distinct from the classical nuclear localization signal (NLS)- and the M9-dependent pathways. Vpr import does not appear to require Ran-mediated GTP hydrolysis and persists under conditions of low energy. Competition experiments further suggest that Vpr directly engages the NPC at two discrete sites. These sites appear to form distal components of a common import pathway used by NLS- and M9-containing proteins. Together, our data suggest that Vpr bypasses many of the soluble receptors involved in import of cellular cargoes. Rather, this viral protein appears to directly access the NPC, a property that may help to ensure the capacity of HIV to replicate in nondividing cellular hosts.
Key Words: Vpr nuclear import HIV nuclear pore complex preintegration complex
Abbreviations used in this paper: βgal, βgalactosidase; GST, glutathione-S-transferase; HIV, human immunodeficiency virus; IBB, importin β binding domain; NLS, nuclear localization signal; NPC, nuclear pore complex; PIC, preintegration complex; WGA, wheat germ agglutinin.
Y. Jenkins is supported by a National Institutes of Health postdoctoral fellowship. K. Weis acknowledges support from the Deutsche Forschungsgemeinschaft. We also acknowledge core support from the University of California San Francisco Center for AIDS Research (P30A127763).
Address all correspondence to Warner C. Greene, Gladstone Institute of Virology and Immunology, University of California, San Francisco, CA 94141-9100. Tel.: (415) 695-3800. Fax: (415) 826-1514. E-mail: wgreene{at}gladstone.ucsf.edu

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