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Viral replication is enhanced by an HIV-1 intersubtype recombination-derived Vpu protein.

De Candia C, Espada C, Duette G, Ghiglione Y, Turk G, Salomón H, Carobene M - Virol. J. (2010)

Bottom Line: Our results showed that BF recombinant Vpu was associated to an increased viral particles production when compared to WT B variant in tetherin-expressing cell lines.The results showed that the replication of the BF Vpu-harboring variant was more efficient in cell cultures than subtype B, reaching a higher frequency in the viral population in a short period of time.This study showed that as a result of intersubtype recombination, a structurally re-organized HIV-1 Vpu has an improved in vitro capacity of enhancing viral replication, and provides evidence of the changes occurring in this protein function that could play an important role in the successful spread of intersubtype recombinant variants.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Reference Center for AIDS, Department of Microbiology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina.

ABSTRACT

Background: Multiple HIV-1 intersubtype recombinants have been identified in human populations. Previous studies from our lab group have shown that the epidemic in Argentina is characterized by the high prevalence of a circulating recombinant form, CRF12_BF, and many related BF recombinant forms. In these genomic structures a recombination breakpoint frequently involved the vpu coding region. Due to the scarce knowledge of Vpu participation in the virion release process and its impact on pathogenesis and of the functional capacities of intersubtype recombinant Vpu proteins, the aim of this work was to perform a comparative analysis on virion release capacity and relative replication capacity among viral variants harboring either a BF recombinant Vpu or a subtype B Vpu.

Results: Our results showed that BF recombinant Vpu was associated to an increased viral particles production when compared to WT B variant in tetherin-expressing cell lines. This observation was tested in the context of a competition assay between the above mentioned variants. The results showed that the replication of the BF Vpu-harboring variant was more efficient in cell cultures than subtype B, reaching a higher frequency in the viral population in a short period of time.

Conclusion: This study showed that as a result of intersubtype recombination, a structurally re-organized HIV-1 Vpu has an improved in vitro capacity of enhancing viral replication, and provides evidence of the changes occurring in this protein function that could play an important role in the successful spread of intersubtype recombinant variants.

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Vpu expression was assessed by specific RT-PCR of vpu mRNA after transfection of HeLa cells. Both untransfected (UT) cells and cells transfected with empty vector (NC) were used as control (right panel). Amplification of β-actin mRNA was also evaluated as an internal control (left panel).
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Figure 2: Vpu expression was assessed by specific RT-PCR of vpu mRNA after transfection of HeLa cells. Both untransfected (UT) cells and cells transfected with empty vector (NC) were used as control (right panel). Amplification of β-actin mRNA was also evaluated as an internal control (left panel).

Mentions: In order to test the vpu mRNA expression, HeLa cell cultures were transfected either with pNL4-3 VpuBF, pNL4-3 VpuB and pNL4-3 ΔVpu plasmids. Forty eight hours post-transfection total mRNA content was obtained from each transfected culture, and vpu mRNA production was evaluated by qualitative RT-PCR. β-actin mRNA was used as control. All transfected cell cultures exhibited comparable expression levels of vpu mRNA (Figure 2).


Viral replication is enhanced by an HIV-1 intersubtype recombination-derived Vpu protein.

De Candia C, Espada C, Duette G, Ghiglione Y, Turk G, Salomón H, Carobene M - Virol. J. (2010)

Vpu expression was assessed by specific RT-PCR of vpu mRNA after transfection of HeLa cells. Both untransfected (UT) cells and cells transfected with empty vector (NC) were used as control (right panel). Amplification of β-actin mRNA was also evaluated as an internal control (left panel).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2967538&req=5

Figure 2: Vpu expression was assessed by specific RT-PCR of vpu mRNA after transfection of HeLa cells. Both untransfected (UT) cells and cells transfected with empty vector (NC) were used as control (right panel). Amplification of β-actin mRNA was also evaluated as an internal control (left panel).
Mentions: In order to test the vpu mRNA expression, HeLa cell cultures were transfected either with pNL4-3 VpuBF, pNL4-3 VpuB and pNL4-3 ΔVpu plasmids. Forty eight hours post-transfection total mRNA content was obtained from each transfected culture, and vpu mRNA production was evaluated by qualitative RT-PCR. β-actin mRNA was used as control. All transfected cell cultures exhibited comparable expression levels of vpu mRNA (Figure 2).

Bottom Line: Our results showed that BF recombinant Vpu was associated to an increased viral particles production when compared to WT B variant in tetherin-expressing cell lines.The results showed that the replication of the BF Vpu-harboring variant was more efficient in cell cultures than subtype B, reaching a higher frequency in the viral population in a short period of time.This study showed that as a result of intersubtype recombination, a structurally re-organized HIV-1 Vpu has an improved in vitro capacity of enhancing viral replication, and provides evidence of the changes occurring in this protein function that could play an important role in the successful spread of intersubtype recombinant variants.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Reference Center for AIDS, Department of Microbiology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentina.

ABSTRACT

Background: Multiple HIV-1 intersubtype recombinants have been identified in human populations. Previous studies from our lab group have shown that the epidemic in Argentina is characterized by the high prevalence of a circulating recombinant form, CRF12_BF, and many related BF recombinant forms. In these genomic structures a recombination breakpoint frequently involved the vpu coding region. Due to the scarce knowledge of Vpu participation in the virion release process and its impact on pathogenesis and of the functional capacities of intersubtype recombinant Vpu proteins, the aim of this work was to perform a comparative analysis on virion release capacity and relative replication capacity among viral variants harboring either a BF recombinant Vpu or a subtype B Vpu.

Results: Our results showed that BF recombinant Vpu was associated to an increased viral particles production when compared to WT B variant in tetherin-expressing cell lines. This observation was tested in the context of a competition assay between the above mentioned variants. The results showed that the replication of the BF Vpu-harboring variant was more efficient in cell cultures than subtype B, reaching a higher frequency in the viral population in a short period of time.

Conclusion: This study showed that as a result of intersubtype recombination, a structurally re-organized HIV-1 Vpu has an improved in vitro capacity of enhancing viral replication, and provides evidence of the changes occurring in this protein function that could play an important role in the successful spread of intersubtype recombinant variants.

Show MeSH
Related in: MedlinePlus