Efficient Particle Production by Minimal Gag Constructs Which Retain the Carboxy-Terminal Domain of Human Immunodeficiency Virus Type 1 Capsid-p2 and a Late Assembly Domain

doi:10.1128/JVI.74.12.5395-5402.2000

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Journal of Virology, June 2000, p. 5395-5402, Vol. 74, No. 12
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Efficient Particle Production by Minimal Gag Constructs Which Retain the Carboxy-Terminal Domain of Human Immunodeficiency Virus Type 1 Capsid-p2 and a Late Assembly Domain

Molly A. Accola,¹^,² Bettina Strack,¹ and Heinrich G. Göttlinger¹^,³^,^*

Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute,¹ and Program in Immunology² and Department of Pathology,³ Harvard Medical School, Boston, Massachusetts 02115

Received 14 September 1999/Accepted 16 March 2000

The human immunodeficiency virus type 1 (HIV-1) Gag precursor Pr55^gag by itself is capable of assembling into retrovirus-like particles(VLP). In the present study, we attempted to identify the minimalGag sequences required for the formation of VLP. Our results showthat about 80% of Pr55^gag can be either deleted or replaced by heterologous sequences withoutsignificantly compromising VLP production. The smallest chimericmolecule still able to efficiently form VLP was only about 16kDa. This minimal Gag construct contained the leucine zipper domainof the yeast transcription factor GCN4 to substitute for the assemblyfunction of nucleocapsid (NC), followed by a P-P-P-P-Y motif toprovide late budding (L) domain function, and retained only themyristylation signal and the C-terminal capsid-p2 domain of Pr55^gag. We also show that the L domain function of HIV-1 p6^gag is not dependent on the presence of an active viral proteaseand that the NC domain of Pr55^gag is dispensable for the incorporation of Vpr intoVLP.

^* Corresponding author. Mailing address: Dana-Farber Cancer Institute, 44 Binney St., Boston, MA 02115. Phone: (617) 632-3067.Fax: (617) 632-3113. E-mail: Heinrich_Gottlinger{at}DFCI.harvard.edu.

Journal of Virology, June 2000, p. 5395-5402, Vol. 74, No. 12
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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