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Journal of Virology, October 2000, p. 9727-9731, Vol. 74, No. 20
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Human Immunodeficiency Virus Type 1 Vpr Protein Is Incorporated into the Virion in Significantly Smaller Amounts than Gag and Is Phosphorylated in Infected Cells

Barbara Müller,1,* Uwe Tessmer,1 Ulrich Schubert,1,2 and Hans-Georg Kräusslich1,dagger

Heinrich-Pette-Institut, D-20251 Hamburg, Germany,1 and Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 0892-04602

Received 7 March 2000/Accepted 18 April 2000

Viral protein R (Vpr) of human immunodeficiency virus type 1 (HIV-1) is a small accessory protein involved in the nuclear import of viral DNA and the growth arrest of host cells. Several studies have demonstrated that a significant amount of Vpr is incorporated into the virus particle via interaction with the p6 domain of Gag, and it is generally assumed that Vpr is packaged in equimolar ratio to Gag. We have quantitated the relative amount of Vpr in purified virions following [35S]cysteine labeling of infected MT-4 cells, as well as by quantitative immunoblotting and found that Vpr is present in a molar ratio of approximately 1:7 compared to capsid. Analysis of isolated core particles showed that Vpr is associated with the mature viral core, despite quantitative loss of p6 from core preparations. Metabolic labeling of infected cells with ortho[32P]phosphate revealed that a small fraction of Vpr is phosphorylated in virions and infected cells.

* Corresponding author. Present address: Abteilung Virologie, Universität Heidelberg, Im Neuenheimer Feld 324, D-69120 Heidelberg, Germany. Phone: 49-6221-565002. Fax: 49-6221-565003. E-mail:Barbara_Mueller{at}med.uni-heidelberg.de.

dagger Present address: Abteilung Virologie, Universität Heidelberg, D-69120 Heidelberg, Germany.

Journal of Virology, October 2000, p. 9727-9731, Vol. 74, No. 20
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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