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Journal of Virology, March 1999, p. 2270-2279, Vol. 73, No. 3
ABL-Basic Research Program, NCI-Frederick
Cancer Research and Development Center, Frederick, Maryland 21702
Received 5 October 1998/Accepted 8 December 1998
Human immunodeficiency virus type 1 (HIV-1) normally assembles into
particles of 100 to 120 nm in diameter by budding through the plasma
membrane of the cell. The Gag polyprotein is the only viral protein
that is required for the formation of these particles. We have used an
in vitro assembly system to examine the assembly properties of
purified, recombinant HIV-1 Gag protein and of Gag missing the
C-terminal p6 domain (Gag
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
In Vitro Assembly Properties of Human
Immunodeficiency Virus Type 1 Gag Protein Lacking the p6
Domain
p6). This system was used previously to
show that the CA-NC fragment of HIV-1 Gag assembled into cylindrical
particles. We now report that both HIV-1 Gag and Gag
p6 assemble
into small, 25- to 30-nm-diameter spherical particles in vitro. The
multimerization of Gag
p6 into units larger than dimers and the
formation of spherical particles required nucleic acid. Removal of the
nucleic acid with NaCl or nucleases resulted in the disruption of the
multimerized complexes. We conclude from these results that (i)
N-terminal extension of HIV-1 CA-NC to include the MA domain results in
the formation of spherical, rather than cylindrical, particles; (ii)
nucleic acid is required for the assembly and maintenance of HIV-1 Gag
p6 virus-like particles in vitro and possibly in vivo; (iii) a wide
variety of RNAs or even short DNA oligonucleotides will support
assembly; (iv) protein-protein interactions within the particle must be
relatively weak; and (v) recombinant HIV-1 Gag
p6 and nucleic acid
are not sufficient for the formation of normal-sized particles.
*
Corresponding author. Mailing address: ABL-Basic
Research Program, NCI-Frederick Cancer Research and Development Center,
P.O. Box B, Frederick, MD 21702. Phone: (301) 846-1844. Fax: (301) 846-7146. E-mail: campbells{at}mail.ncifcrf.gov.
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