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Journal of Virology, March 1999, p. 1931-1940, Vol. 73, No. 3
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Actin Associates with the Nucleocapsid Domain of the Human Immunodeficiency Virus Gag Polyprotein

Thomas Wilk, Brent Gowen,dagger and Stephen D. Fuller*

Structural Biology Programme, European Molecular Biology Laboratory, 69117 Heidelberg, Germany

Received 1 April 1998/Accepted 7 December 1998

Recently, it was shown that actin molecules are present in human immunodeficiency virus type 1 (HIV-1) particles. We have examined the basis for incorporation and the location of actin molecules within HIV-1 and murine retrovirus particles. Our results show that the retroviral Gag polyprotein is sufficient for actin uptake. Immunolabeling studies demonstrate that actin molecules localize to a specific radial position within the immature particle, clearly displaced from the matrix domain underneath the viral membrane but in proximity to the nucleocapsid (NC) domain of the Gag polyprotein. When virus or subviral Gag particles were disrupted with nonionic detergent, actin molecules remained associated with the disrupted particles. Actin molecules remained in a stable complex with the NC cleavage product (or an NC-RNA complex) after treatment of the disrupted HIV-1 particles with recombinant HIV-1 protease. In contrast, matrix and capsid molecules were released. The same result was obtained when mature HIV-1 particles were disrupted with detergent. Taken together, these results indicate that actin molecules are associated with the NC domain of the viral polyprotein.

* Corresponding author. Mailing address: Structural Biology Programme, European Molecular Biology Laboratory, Postfach 10.2209, 69117 Heidelberg, Germany. Phone: 49-6221-387-265. Fax: 49-6221-387-306. E-mail: Fuller{at}EMBL-Heidelberg.DE.

dagger Present address: Cryo-TEM Facility, Department of Biochemistry, Imperial College of Science, Technology, and Medicine, London, United Kingdom SW7 2AZ.

Journal of Virology, March 1999, p. 1931-1940, Vol. 73, No. 3
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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