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Journal of Virology, February 2008, p. 1229-1237, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.00540-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Hsp40 Facilitates Nuclear Import of the Human Immunodeficiency Virus Type 2 Vpx-Mediated Preintegration Complex{triangledown}

Xiaogang Cheng,1 Michael Belshan,2 and Lee Ratner1*

Division of Molecular Oncology, Departments of Medicine and Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110,1 Department of Medical Microbiology and Immunology, Creighton University School of Medicine, Omaha, Nebraska 681782

Received 14 March 2007/ Accepted 9 November 2007

Human immunodeficiency virus type 2 (HIV-2) Vpx is required for nuclear translocation of the viral preintegration complex (PIC) in quiescent cells. In order to decipher the mechanism of action of Vpx, a cDNA library was screened with the yeast two-hybrid assay, resulting in the identification of heat shock protein 40, Hsp40/DnaJB6, as a Vpx-interactive protein. Interaction with Vpx was confirmed by glutathione S-transferase (GST) pull-down and coimmunoprecipitation assays. Overexpression of Hsp40/DnaJB6 enhanced Vpx nuclear import, whereas overexpression of a nuclear localization mutant of Hsp40/DnaJB6 (H31Q) or down-regulation of Hsp40/DnaJB6 by small interfering RNA (siRNA) reduced the nuclear import of Vpx. Hsp40/DnaJB6 competed with the Pr55Gag precursor protein for the binding of Vpx and incorporation into virus-like particles. Overexpression of Hsp40/DnaJB6 promoted viral PIC nuclear import, whereas siRNA down-regulation of Hsp40/DnaJB6 inhibited PIC nuclear import. These results demonstrate a role for Hsp40/DnaJB6 in the regulation of HIV-2 PIC nuclear transport.

* Corresponding author. Mailing address: Box 8069, 660 S. Euclid Ave., Washington University School of Medicine, St. Louis, MO 63110. Phone: (314) 362-8836. Fax: (314) 747-2120. E-mail: lratner{at}im.wustl.edu

{triangledown} Published ahead of print on 21 November 2007.

Journal of Virology, February 2008, p. 1229-1237, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.00540-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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