Requirements for incorporation of Pr160gag-pol from human immunodeficiency virus type 1 into virus-like particles.
The roles of the human immunodeficiency virus precursor polyproteins Pr55gag and Pr160gag-pol in viral core assembly were studied in CMT3-COS cells. To do this, the precursors were expressed separately by using a simian virus 40 late replacement vector system described previously. Consistent with previously published data, our results show that the Pr55gag precursor, when expressed alone, was able to form particles which had an immature morphology and that particle formation required the presence of a myristate addition signal at the amino terminus of the precursor. In contrast, the Pr160gag-pol precursor was not able to form particles when expressed alone, although it still underwent proteolytic processing. Coexpression of the two precursor polyproteins from separate vectors in the same cell resulted in processing of the Pr55gag in trans by the protease embedded in Pr160gag-pol and the formation of virus-like particles containing the products of both precursors. Proteolytic processing occurred independently of the presence of a functional myristate addition signal on either precursor. On the other hand, removal of myristate from one or the other precursor had nonreciprocal effects on virus particle formation. Cells expressing Pr55gag lacking myristate and Pr160gag-pol containing it did not produce particles. Cells expressing a myristylated Pr55gag and unmyristylated Pr160gag-pol still produced virus-like particles which contained nearly normal amounts of Pr160gag-pol. The results suggest that the incorporation of Pr160gag-pol into particles is largely determined by intermolecular protein-protein interactions between the two precursor polypeptides.
Smith
A J
AJ
Department of Microbiology, State University of New York, Buffalo 14214.
Srinivasakumar
N
N
Hammarskjöld
M L
ML
Rekosh
D
D
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015683:Gene Products, gag / Q000378:metabolism*; D015684:Gene Products, pol / Q000378:metabolism*; D015497:HIV-1 / Q000254:growth & development* / Q000648:ultrastructure; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009024:Morphogenesis; D009226:Myristates / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D014162:Transfection
D015683:Gene Products, gag; D015684:Gene Products, pol; D009226:Myristates; D011498:Protein Precursors; C059767:p55 gag precursor protein, Human immunodeficiency virus 1