AEGiS-12IAC: Immunocytochemical localizations of HIV-1 Gag proteins and formation of O-shaped virion particles associate with p17.

12th International AIDS Conference


Geneva, Switzerland - June 28-July 3, 1998

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Immunocytochemical localizations of HIV-1 Gag proteins and formation of O-shaped virion particles associate with p17.

Int Conf AIDS 1998 Jun 28-Jul 3; 12:10 (abstract no. 11138)

Wang JJ, Chiou CT, Chiang PH, Hortor B, Ratner L
Inst Biol & Anat, Natl Def Med Ctr, Taipei, ROC.

OBJECTIVES: To identify the components and localizations of viral proteins during HIV-1 assembly.

DESIGN: Transfection of Gag protein p55G1 alone can form virion particles. Different constructs of Gag/Pol proteins were designed and expressed in human T cell lines. Intracellular trafficking of the components of virion particles by a modified immunocytochemical technique was identified.

METHODS: Vaccinia virus with pTM3 vector infection/transfection system were performed with recombined Gag/Pol proteins in Jurkat cells, 293T cells and BSC40 cells for various time periods. Immunocytochemical data were analyzed with confocal and immunoelectron microscopy.

RESULTS: High titer of antigenicity of p24 and p17 in the well-fixed and regularly embedded samples were preserved for the immuno-gold labeling. Budding of O-shaped virion particles on the cell membrane was found in p55G1 transfections and labeled by p24-Ab conjugated with gold particles. Assembly of p24 occurred in the cytosol on free ribosomal granules, in rER and mitochondrial membranes but not in the Golgi apparatus. Accumulation of virion particles at the perinuclear pores indicated that something from the nuclear may joining the particles. C-shaped free virion particles were found accumulated on the vacuolar membrane and in the cytosol but not on the cell membrane when transfected with p39A1, p41A1 and p55A1 omitting whole or part of matrix protein, p17.

CONCLUSIONS: Under the vaccinia infection/transfection system, p55G1 particles are assembled among free ribosomes or on membranous structures. The matrix protein is essential in directing Gag protein to the cell membrane and also necessary for forming an O-shaped virion particle.

Keywords: AEGIS, Gene Products, gag, Virion, HIV-1, Gene Products, pol, Vaccinia virus, Vaccinia, Viral Proteins, Transfection, Jurkat Cells, Human, genetics, ICA12KWDaegis,geneproducts,gag,virion,hiv-1,geneproducts,pol,vacciniavirus,vaccinia,viralproteins,transfection,jurkatcells,human,genetics,ica12
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