CHARACTERIZATION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PR55(GAG) MEMBRANE ASSOCIATION IN A CELL-FREE SYSTEM - REQUIREMENT FOR A C-TERMINAL DOMAIN

被引:40
作者
PLATT, EJ [1 ]
HAFFAR, OK [1 ]
机构
[1] BRISTOL MYERS SQUIBB PHARMACEUT RES INST, SEATTLE, WA 98121 USA
关键词
D O I
10.1073/pnas.91.10.4594
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Association of the human immunodeficiency virus type 1 (HIV-1) gag polyprotein precursor with cellular membranes is necessary for assembly of virions. We used in vitro synthesized HIV-1 gag to study its association with isolated cellular membranes. Rabbit reticulocyte lysates programmed with HIV-1 gag mRNA incorporated [S-35]methionine and [H-3]myristate into two predominant species of 55 kDa and 40 kDa. Radioimmunoprecipitation with HIV-1-specific antibodies suggested that the 55-kDa protein represented the polyprotein precursor (Pr55(gag)), while the 40-kDa protein was a mixture of N- or C-terminal truncations of the gag precursor. The Pr55(gag) protein bound to cellular membranes, while the 40-kDa mixed protein species did not. Membrane binding studies with C terminus-truncated and point mutants revealed that the seven-amino acid sequence located between the two Cys-His arrays in the nucleocapsid region was necessary for stable association to occur. Therefore, we propose that signals in addition to myristate are required for the membrane association of HIV-1 gag proteins and that these signals include a domain in the nucleocapsid protein.
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页码:4594 / 4598
页数:5
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