Interaction and co-encapsidation of human immunodeficiency virus type 1 Gag and Vif recombinant proteins.

Huvent, Isabelle; Spire, Bruno; Carri√®re, C; Tournier, J; Gay, Bernard; Fournier, C; Vigne, Robert; Courcoul, Marianne; Boulanger, Pierre; Hong, Saw See

doi:10.1099/0022-1317-79-5-1069

Cited by 57 publications

(119 citation statements)

References 44 publications

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“…Although immunofluorescence studies showed a co-localization of Gag and Vif in the cell (38), co-sedimentation studies indicated an interaction of Vif only with some early viral assembly intermediates, and the presence of Vif in mature virions remains controversial (39 -46). In insect cells infected with baculovirus expressing Gag and Vif, it was estimated that there were 70 Vif molecules per 2000 Gag molecules in extracellular Gag particles or one molecule of Vif for every 30 molecules of Gag (47). If single Gag molecules bound to Vif at this same ratio within an APOBEC3G/Vif/Gag complex destined for degradation in the proteosome, this would account for only 3.5% of Gag molecules produced, and a change in Gag distribution in the cell would not be detectable by our Western blot assay.…”

Section: Discussionmentioning

confidence: 99%

The Interaction between HIV-1 Gag and APOBEC3G

Cen

Guo

Niu

et al. 2004

Journal of Biological Chemistry

235

238

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APOBEC3G, a member of an RNA/DNA cytidine deaminase superfamily, has been identified as a cellular inhibitor of HIV-1 infectivity, possibly through the dC to dU deamination of the first minus strand cDNA synthesized during reverse transcription. Virions incorporate APOBEC3G during viral assembly in non-permissive cells, and this incorporation is inhibited by the viral protein Vif. The mechanism of APOBEC3G incorporation into HIV-1 is examined in this report. In the absence of Vif, cytoplasmic APOBEC3G becomes membranebound in cells expressing HIV-1 Gag, and its incorporation into Gag viral-like particles (VLPs) is proportional to the amount of APOBEC3G expressed in the cell. The expression of Vif, or mutant Gag unable to bind to membrane, prevents the APOBEC3G association with membrane. HIV-1 Gag alone among viral proteins is sufficient for packaging of APOBEC3G into Gag VLPs, and this incorporation requires the presence of Gag nucleocapsid. The presence of amino acids 104 -156 in APOBEC3G, located in the linker region between two zinc coordination motifs, is also required for its incorporation into Gag VLPs. Evidence against an RNA bridge facilitating the Gag/APOBEC3G interaction includes data indicating that 1) the incorporation of APOBEC3G occurs independently of viral genomic RNA, 2) a Gag/APOBEC3G complex is immunoprecipitated from cell lysate after RNase treatment, and 3) the zinc coordination motif, rather than the regions flanking this motif, have been implicated in RNA binding in another family member, APOBEC1.

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Section: Discussionmentioning

confidence: 99%

The Interaction between HIV-1 Gag and APOBEC3G

Cen

Guo

Niu

et al. 2004

Journal of Biological Chemistry

235

238

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“…In certain cases, polyclonal anti-GAG antibody (laboratory-made; see Ref. 25) was used. Antiserum against HEED was prepared in rabbit, by injection of purified HEEDNt120 -535 protein obtained by affinity purification of GST-fused protein as described above.…”

Section: Gel Electrophoresis and Immunological Analysesmentioning

confidence: 99%

“…38) was used for biopanning onto immobilized protein ligate. Specific ligand elution of phages has been described in previous studies (25,39,40). Purified MA143 was immobilized on enzyme-linked immunosorbent assay plates used as the solid support.…”

Section: Phage-displayed Hexapeptide Library and Biopanningmentioning

confidence: 99%

“…Cell specimens were included, sectioned, and processed for conventional EM or IEM, according to previously described methods (21,25). Mouse monoclonal anti-MA antibody and rabbit anti-HEED antibody were used for IEM, with the corresponding colloidal gold-labeled complementary antibodies, 5-nm gold-tagged anti-mouse Ig antibody, and 10-nm gold-tagged anti-rabbit Ig antibody, respectively.…”

Section: Electron and Immunoelectron Microscopy (Em And Iem)mentioning

confidence: 99%

“…We have generated a variety of mutations in the gag gene of HIV-1, consisting of insertions, substitutions, or deletions in the different structural domains of the Pr55GAG and expressed the corresponding protein mutants in recombinant baculovirus-infected insect cells (21)(22)(23)(24)(25)(26)(27)(28). One of our C-truncated GAG mutants, carrying an amber mutation at codon 143 in the N-terminal portion of the capsid (CA) domain (GAGamb143; see Ref.…”

mentioning

confidence: 99%

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HEED, the Product of the Human Homolog of the Murineeed Gene, Binds to the Matrix Protein of HIV-1

Peytavi

Hong

d’Angeac

et al. 1999

Journal of Biological Chemistry

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heed, the human homolog of mouse eed and Drosophila esc, two members of the trithorax (trx) and Polycomb group (Pc-G) of genes, was isolated by screening an activated lymphocyte cDNA library versus the immunodeficiency virus type 1 (HIV-1) MA protein used as a bait in a two-hybrid system in yeast. The human EED protein (HEED) had 99.5% identity with the mouse EED protein and contained seven WD repeats. Two heed gene transcripts were identified, with a putative 407-nucleotidelong intron, giving rise to two HEED protein isoforms of 535 and 494 residues in length, respectively. The shorter HEED isoform, originated from the unspliced message, lacked the seventh WD repeat.

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Morphopoietic Determinants of HIV-1 Gag Particles Assembled in Baculovirus-Infected Cells

et al. 1998

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The determinants for HIV-1 particle morphology were investigated using various deletion and insertion mutants of the Gap precursor protein (Gag) expressed in baculovirus-infected cells and ultrastructural analysis of membrane-enveloped Gag particles under the electron microscope. Five discrete regions were found to influence the size, the variability in dimension, and the sphericity of the particles: (i) the matrix (MA) N-terminal domain, within residues 10-21, the junctions of (ii) MA-CA (capsid), (iii) CA-spacer peptide SP1 and (iv) nucleocapsid (NC)-SP2, and (v) the p6gag C-terminus. Internal regions (ii), (iii), and (iv) contained HIV-1 protease cleavage sites separating major structural domains. No particle assembly was observed for am276, a MA-CA polyprotein mutant lacking the C-terminal third of the CA domain. However, MA-CA domains including the MHR (residues 277-306), or downstream sequence to CA residue 357, resulted in the assembly into tubular or filamentous structures, suggesting a helical symmetry of Gag packing. Mutant amb374, derived from amb 357 by further addition of the heptadecapeptide motif HKARVLAEAMSQVTNSA, overlapping the CA-SP1 junction and the SP1 domain, showed a drastic change in the pattern of Gag assembly, compared to amb357, with formation of spherical particles. These data suggested a novel function for the spacer domain SP1, acting as a spherical shape determinant of the Gag particle which would negatively affect the helical symmetry of assembly of the Gag precursor molecules conferred by the MHR and the downstream CA sequence, within residues 307-357.

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Interaction and co-encapsidation of human immunodeficiency virus type 1 Gag and Vif recombinant proteins.

Abstract: Human immunodeficiency virus type 1 (HIV-1

Cited by 57 publications

References 44 publications

The Interaction between HIV-1 Gag and APOBEC3G

The Interaction between HIV-1 Gag and APOBEC3G

HEED, the Product of the Human Homolog of the Murineeed Gene, Binds to the Matrix Protein of HIV-1

Morphopoietic Determinants of HIV-1 Gag Particles Assembled in Baculovirus-Infected Cells

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