Vif is crucial for human immunodeficiency virus type 1 proviral DNA synthesis in infected cells

Schwedler, Uta von; Song, Jinping; Aiken, Christopher; Trono, Didier

doi:10.1128/jvi.67.8.4945-4955.1993

Cited by 410 publications

(287 citation statements)

References 44 publications

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“…All viruses used in this study were transiently expressed by calcium phosphate transfection of 40 μg of each proviral DNA construct as described previously (von Schwedler et al, 1993). It is important to emphasize that wild-type and mutant viruses used in the present study were all produced from 293T cells, harvested at the same time and purified under the same conditions.…”

Section: Methodsmentioning

confidence: 99%

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Host cyclophilin A mediates HIV-1 attachment to target cells via heparans

1999

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The present study proposes a novel mode of action for cyclophilin A (CypA) in the HIV-1 life cycle. We demonstrate that CypA-deficient viruses do not replicate because they fail to attach to target cells. We show that CypA is exposed at the viral membrane and mediates HIV-1 attachment. We identify heparan as the exclusive cellular binding partner for CypA. Furthermore, CypA binds directly to heparan via a domain rich in basic residues similar to known heparinbinding motifs. This interaction between exposed CypA and cell surface heparans represents the initial step of HIV-1 attachment and is a necessary precursor to gp120-binding to CD4. In conclusion, HIV-1 attachment to target cells is a multi-step process that requires an initial CypA-heparan interaction followed by the gp120-CD4 interaction.

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Section: Methodsmentioning

confidence: 99%

“…Purification and immunoblot analysis of viruses produced from 293T transfected cells were conducted as previously described (von Schwedler et al, 1993). Anti-MA and anti-gp41 IgG were purchased from Advanced Biotechnologies International.…”

Section: Methodsmentioning

confidence: 99%

Host cyclophilin A mediates HIV-1 attachment to target cells via heparans

1999

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“…HIV-1 viruses encoding all but the Vif protein were produced from the pR9⌬vif plasmid (60). The plasmids required for the LINE-1-driven Alu retrotransposition assay were a kind gift from J. V. Moran (pALU_pAϩ_neoTET and pJM101_L1.RP_NoNeo) (19,29).…”

Section: Constructsmentioning

confidence: 99%

Functional Analysis and Structural Modeling of Human APOBEC3G Reveal the Role of Evolutionarily Conserved Elements in the Inhibition of Human Immunodeficiency Virus Type 1 Infection and Alu Transposition

Bulliard

Turelli

Röhrig

et al. 2009

J Virol

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Retroelements are important evolutionary forces but can be deleterious if left uncontrolled. Members of the human APOBEC3 family of cytidine deaminases can inhibit a wide range of endogenous, as well as exogenous, retroelements. These enzymes are structurally organized in one or two domains comprising a zinc-coordinating motif. APOBEC3G contains two such domains, only the C terminal of which is endowed with editing activity, while its N-terminal counterpart binds RNA, promotes homo-oligomerization, and is necessary for packaging into human immunodeficiency virus type 1 (HIV-1) virions. Here, we performed a large-scale mutagenesisbased analysis of the APOBEC3G N terminus, testing mutants for (i) inhibition of vif-defective HIV-1 infection and Alu retrotransposition, (ii) RNA binding, and (iii) oligomerization. Furthermore, in the absence of structural information on this domain, we used homology modeling to examine the positions of functionally important residues and of residues found to be under positive selection by phylogenetic analyses of primate APOBEC3G genes. Our results reveal the importance of a predicted RNA binding dimerization interface both for packaging into HIV-1 virions and inhibition of both HIV-1 infection and Alu transposition. We further found that the HIV-1-blocking activity of APOBEC3G N-terminal mutants defective for packaging can be almost entirely rescued if their virion incorporation is forced by fusion with Vpr, indicating that the corresponding region of APOBEC3G plays little role in other aspects of its action against this pathogen. Interestingly, residues forming the APOBEC3G dimer interface are highly conserved, contrasting with the rapid evolution of two neighboring surface-exposed amino acid patches, one targeted by the Vif protein of primate lentiviruses and the other of yet-undefined function.

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“…The dilution resulting in less then 30% GFP-positive cells was used to calculate the number of TU per mL. The particle titer for both the LV-CMV-GFP and the LV-CMV-GDNF vectors was estimated by an RNA slot blot hybridization technique (von Schwedler et al, 1993). The ratio between the number of TU and the particle titer for the LV-CMV-GFP vector was used to estimate the number of TU for the LV-CMV-GDNF vector.…”

Section: Production Of Lentiviral Vectorsmentioning

confidence: 99%

Ex vivo gene delivery of GDNF using primary astrocytes transduced with a lentiviral vector provides neuroprotection in a rat model of Parkinson's disease

Ericson

Georgievska²,

Lundberg³

2005

Eur J of Neuroscience

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Astrocytes are, as normal constituents of the brain, promising vehicles for ex vivo gene delivery to the central nervous system. In the present study, we have used a lentiviral vector encoding glial cell line-derived neurotrophic factor (GDNF) to transduce rat-derived primary astrocytes, in order to evaluate their potential for long-term transgene expression in vivo and neuroprotection in a rat model of Parkinson's disease. Following transplantation of GDNF-transduced astrocytes to the intact striatum, the level of released GDNF was 2.93 +/- 0.28 ng/mg tissue at 1 week post-grafting, reduced to 0.42 +/- 0.12 ng/mg tissue at 4 weeks, and thereafter was maintained at this level throughout the experiment (12 weeks; 0.53 +/- 0.068 ng/mg tissue). Similarly, grafting to the substantia nigra (SN) resulted in a significant overexpression of GDNF ( approximately 0.20 ng/mg tissue) at 1 week. Intact animals receiving transplants of GDNF-transduced astrocytes displayed an increased contralateral turning (5.39 +/- 1.19 turns/min) in the amphetamine-induced rotation test, which significantly correlated with the GDNF tissue levels measured in the striatum, indicating a stimulatory effect of GDNF on the dopaminergic function. Transplantation of GDNF-transduced astrocytes to the SN 1 week prior to an intrastriatal 6-hydroxydopamine lesion provided a significant protection of nigral tyrosine hydroxylase-positive cells. By contrast, when the cells were transplanted to the striatum, the level of released GDNF was not sufficient to rescue the striatal fibers and, hence, to protect the nigral dopaminergic neurons. Overall, our results suggest that genetically modified astrocytes expressing GDNF can provide neuroprotection in a rat model of Parkinson's disease following transplantation to the SN.

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Vif is crucial for human immunodeficiency virus type 1 proviral DNA synthesis in infected cells

Cited by 410 publications

References 44 publications

Host cyclophilin A mediates HIV-1 attachment to target cells via heparans

Host cyclophilin A mediates HIV-1 attachment to target cells via heparans

Functional Analysis and Structural Modeling of Human APOBEC3G Reveal the Role of Evolutionarily Conserved Elements in the Inhibition of Human Immunodeficiency Virus Type 1 Infection and Alu Transposition

Ex vivo gene delivery of GDNF using primary astrocytes transduced with a lentiviral vector provides neuroprotection in a rat model of Parkinson's disease

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