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Journal of Virology, October 1998, p. 7950-7959, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Analysis of Minimal Human Immunodeficiency Virus Type 1 gag Coding Sequences Capable of Virus-Like Particle Assembly and Release

Chin-Tien Wang,* Hsiu-Yu Lai, and Jue-Jyh Li

Institute of Clinical Medicine, National Yang-Ming University, and Department of Medical Research, Veterans General Hospital-Taipei, Taipei, Taiwan 11217, Republic of China

Received 16 January 1998/Accepted 15 June 1998

We have constructed a series of human immunodeficiency virus (HIV) gag mutants by progressive truncation of the gag coding sequence from the C terminus and have combined these mutants with an assembly-competent matrix domain deletion mutation (Delta MA). By using several methods, the particle-producing capabilities of each mutant were examined. Our analysis indicated that truncated Gag precursors lacking most of C-terminal gag gene products assembled and were released from 293T cells. Additionally, a mutant with a combined deletion of the MA (Delta MA) and p6 domains even produced particles at levels comparable to that of the wild-type (wt) virus. However, most mutants derived from combination of the Delta MA and the C-terminal truncation mutations did not release particles as well as the wt. Our smallest HIV gag gene product capable of virus-like particle formation was a 28-kDa protein which consists of a few MA amino acids and the CA-p2 domain. Sucrose density gradient fractionation analysis indicated that most mutants exhibited a wt retrovirus particle density. Exceptions to this rule were mutants with an intact MA domain but deleted downstream of the p2 domains. These C-terminal truncation mutants possessed particle densities of 1.13 to 1.15 g/ml, lower than that of the wt. The N-terminal portions of the CA domain, which have been shown to be dispensable for core assembly, became critical when most of the MA domain was deleted, suggesting a requirement for an intact CA domain to assemble and release particles.

* Corresponding author. Mailing address: Department of Medical Research, Veterans General Hospital-Taipei, No. 201, Sec. 2, Shih-pai Rd., Shih-pai, Taipei, Taiwan 11217. Phone: 886-2-2874-2121, ext. 2655. Fax: 886-2-2874-2279. E-mail: ctwang{at}vghtpe.gov.tw.

Journal of Virology, October 1998, p. 7950-7959, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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