Subcellular localization of the Vif protein of human immunodeficiency virus type 1

Goncalves, J; Jallepalli, Prasad V.; Gabuzda, Dana

doi:10.1128/jvi.68.2.704-712.1994

Cited by 120 publications

(84 citation statements)

References 48 publications

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“…Formation of the viral particles is therefore probably normal, allowing the analysis of the role of vif during the late steps of virus replication. The fact that the Vif protein has been localized to the cytoplasmic side of the cellular membranes (12) supports the results presented in the present study: vif acts at the level of particle formation. This observation raises some additional questions; for example, on which modification and on which viral or cellular target protein does Vif act?…”

Section: Discussionsupporting

confidence: 92%

“…It encodes a 23-kDa protein which is absent from virions but present in infected cells (17). Recent observations have indicated that HIV-1 Vif is located in the cytoplasm both in a cytosolic form and associated with the cytoplasmic side of cellular membranes (12). This protein is encoded by all lentivirus genomes except that of equine infectious anemia virus (19) and is antigenic, since antibodies against Vif are detected in the sera of infected individuals (1,17).…”

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confidence: 99%

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Peripheral blood mononuclear cells produce normal amounts of defective Vif- human immunodeficiency virus type 1 particles which are restricted for the preretrotranscription steps

Courcoul

Patience

Rey

et al. 1995

J Virol

116

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Previous studies have demonstrated the absence of viral replication of Vif ؊ mutants in stimulated primary blood mononuclear cells (PBMC). Human immunodeficiency virus type 1 strain NDK Vif ؊ mutants were propagated on the semipermissive CEM cell line, and the viral stock obtained was compared with the wild-type virus during a single cycle in PBMC. The Vif ؊ virus was able to enter PBMC with the same efficiency as the wild type, as demonstrated by quantification of the strong-stop cDNA, and retrotranscription was observed for both viruses within 4 h postinfection. Using a PCR assay with an Alu-long terminal repeat pair of primers, we detected integration for both the wild-type and Vif ؊ viruses. We then used qualitative and quantitative reverse transcription-mediated PCR techniques to study the steady-state level of intracellular and extracellular viral RNAs. All mRNA species were detected in PBMC infected with the wild-type virus or with the Vif ؊ virus 36 h postinfection. Furthermore, quantification of viral RNA released from infected cells demonstrated similar levels of virus produced after a unique cycle of replication. However, the Vif ؊ virus obtained after one replication cycle in PBMC was unable to initiate retrotranscription in permissive target cells. These data strongly suggest that the failure to infect target cells is due to a defect in the formation of the viral particle in PBMC.

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Section: Discussionsupporting

confidence: 92%

mentioning

confidence: 99%

Peripheral blood mononuclear cells produce normal amounts of defective Vif- human immunodeficiency virus type 1 particles which are restricted for the preretrotranscription steps

Courcoul

Patience

Rey

et al. 1995

J Virol

116

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“…A cocultivation method was used to produce large quantities of wild-type and vif mutant virus in a nonpermissive CEM cell line (11,14). Infection of CEM cells was initiated by cocultivation with COS cells transfected with 5 g of wild-type or vif mutant HXB2 DNA by the DEAE-dextran method (14,18) from 36 to 72 h after transfection. Infection of SupT1 cells was initiated by DEAE-dextran transfection with 10 g of viral DNA (14).…”

Section: Methodsmentioning

confidence: 99%

Role of Vif in human immunodeficiency virus type 1 reverse transcription

Goncalves

Korin

Zack

et al. 1996

J Virol

Self Cite

110

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The Vif protein of human immunodeficiency virus type 1 (HIV-1) is important for virion infectivity. Previous studies have shown that vif mutant HIV-1 virions are defective in their ability to synthesize proviral DNA in vivo. Here, we examine the role of Vif in viral DNA synthesis in the endogenous reverse transcriptase (RT) reaction, an in vitro assay in which virions synthesize viral DNA by using endogenous viral RNA as a template. vif mutant virions showed a significant reduction in endogenous RT activity despite similar levels of exogenous RT activity. Analysis of the viral DNA products on agarose gels demonstrated that this reflects reduced synthesis of short minus-and plus-strand DNA products in addition to those of full genomic length. Quantitative PCR analysis of endogenous reverse transcription provided further evidence for reduced formation of both initial and completed reverse transcripts. Vif had no effect on genomic RNA dimerization or the stability of the RNA dimer linkage. These results suggest that Vif is important for an early event after virus entry but preceding or during the early stages of viral DNA synthesis. This may be due to an intrinsic effect on reverse transcription or a preceding postentry event(s), such as virion uncoating or disassembly of the virion core. Drugs targeted to Vif function may provide a new therapeutic approach to inhibiting HIV-1 reverse transcription.

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“…HAtagged hA3G was probed with a mouse anti-HA (Sigma) monoclonal antibody at a dilution of 1:5000. For HIV-1 Vif detection, a rabbit anti-Vif polyclonal antibody (Goncalves et al, 1994) was used at a 1:2000 dilution. The secondary antibody was a sheep anti-mouse or donkey anti-rabbit horseradish peroxidase-(HRP) conjugated antibody (Invitrogen), both at 1:5000 dilutions.…”

Section: Western Immunoblotmentioning

confidence: 99%

APOBEC3G cytidine deaminase association with coronavirus nucleocapsid protein

Wang

2009

Virology

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We previously reported that replacing HIV-1 nucleocapsid (NC) domain with SARS-CoV nucleocapsid (N) residues 2-213, 215-421, or 234-421 results in efficient virus-like particle (VLP) production at a level comparable to that of wild-type HIV-1. In this study we demonstrate that these chimeras are capable of packaging large amounts of human APOBEC3G (hA3G), and that an HIV-1 Gag chimera containing the carboxyl-terminal half of human coronavirus 229E (HCoV-229E) N as a substitute for NC is capable of directing VLP assembly and efficiently packaging hA3G. When co-expressed with SARS-CoV N and M (membrane) proteins, hA3G was efficiently incorporated into SARS-CoV VLPs. Data from GST pull-down assays suggest that the N sequence involved in N-hA3G interactions is located between residues 86 and 302. Like HIV-1 NC, the SARS-CoV or HCoV-229E N-associated with hA3G depends on the presence of RNA, with the first linker region essential for hA3G packaging into both HIV-1 and SARS-CoV VLPs. The results raise the possibility that hA3G is capable of associating with different species of viral structural proteins through a potentially common, RNA-mediated mechanism.

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Subcellular localization of the Vif protein of human immunodeficiency virus type 1

Abstract: Biochemical characteristics of the association of Vif with cellular membranes and its membrane topology were also examined. These studies also investigate the roles of the N and C termini and highly conserved regions of Vif in membrane targeting.

Cited by 120 publications

References 48 publications

Peripheral blood mononuclear cells produce normal amounts of defective Vif- human immunodeficiency virus type 1 particles which are restricted for the preretrotranscription steps

Peripheral blood mononuclear cells produce normal amounts of defective Vif- human immunodeficiency virus type 1 particles which are restricted for the preretrotranscription steps

Role of Vif in human immunodeficiency virus type 1 reverse transcription

APOBEC3G cytidine deaminase association with coronavirus nucleocapsid protein

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