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NS2 is dispensable for efficient assembly of hepatitis C virus-like particles in a bipartite trans-encapsidation system.

Bentham MJ, Marraiki N, McCormick CJ, Rowlands DJ, Griffin S - J. Gen. Virol. (2014)

Bottom Line: This closely resembled replicon-mediated NS2 trans-complementation, confirming that baculovirus expression of HCV proteins did not unduly affect particle production.Furthermore, this suggests that separation of structural protein expression from replicating HCV RNAs that are destined to be packaged alleviates an early stage requirement for NS2 during particle formation.This highlights our current lack of understanding of how NS2 mediates assembly, yet comparison of full-length and bipartite systems may provide further insight into this process.

View Article: PubMed Central - PubMed

Affiliation: Leeds Institute of Cancer & Pathology (LICAP), and Leeds CRUK Clinical Centre, Faculty of Medicine and Health, St James's University Hospital, University of Leeds, Beckett St., Leeds, West Yorkshire LS9 7TF, UK.

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Characterization of secreted HCV-LP produced in the presence/absence of NS2. Top, infectivity profile of secreted HCV-LP generated in the absence/presence of NS2 (see diagram) following isopycnic density gradient ultracentrifugation in a 10–40 % iodixinol/PBS gradient. Bottom, core immunoblot of methanol-precipitated gradient fractions.
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f5: Characterization of secreted HCV-LP produced in the presence/absence of NS2. Top, infectivity profile of secreted HCV-LP generated in the absence/presence of NS2 (see diagram) following isopycnic density gradient ultracentrifugation in a 10–40 % iodixinol/PBS gradient. Bottom, core immunoblot of methanol-precipitated gradient fractions.

Mentions: Consistent with bulk infectivity measurements (Fig. 4a), levels of secreted infectious HCV-LP produced in the absence of NS2 were much lower than those formed in its presence (Fig. 5, top panel). Nevertheless, core protein was detectable in the peak fractions from gradients of particles formed in the absence of NS2 (Fig. 5, lower panel). However, the reduction in protein levels was not proportionate to the loss of infectivity, suggesting that the specific infectivity of secreted HCV-LP may have been affected. It is also not possible to rule out that HCV-LP present within the secreted compartment result from cell lysis liberating particles that might not otherwise be efficiently secreted.


NS2 is dispensable for efficient assembly of hepatitis C virus-like particles in a bipartite trans-encapsidation system.

Bentham MJ, Marraiki N, McCormick CJ, Rowlands DJ, Griffin S - J. Gen. Virol. (2014)

Characterization of secreted HCV-LP produced in the presence/absence of NS2. Top, infectivity profile of secreted HCV-LP generated in the absence/presence of NS2 (see diagram) following isopycnic density gradient ultracentrifugation in a 10–40 % iodixinol/PBS gradient. Bottom, core immunoblot of methanol-precipitated gradient fractions.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4202265&req=5

f5: Characterization of secreted HCV-LP produced in the presence/absence of NS2. Top, infectivity profile of secreted HCV-LP generated in the absence/presence of NS2 (see diagram) following isopycnic density gradient ultracentrifugation in a 10–40 % iodixinol/PBS gradient. Bottom, core immunoblot of methanol-precipitated gradient fractions.
Mentions: Consistent with bulk infectivity measurements (Fig. 4a), levels of secreted infectious HCV-LP produced in the absence of NS2 were much lower than those formed in its presence (Fig. 5, top panel). Nevertheless, core protein was detectable in the peak fractions from gradients of particles formed in the absence of NS2 (Fig. 5, lower panel). However, the reduction in protein levels was not proportionate to the loss of infectivity, suggesting that the specific infectivity of secreted HCV-LP may have been affected. It is also not possible to rule out that HCV-LP present within the secreted compartment result from cell lysis liberating particles that might not otherwise be efficiently secreted.

Bottom Line: This closely resembled replicon-mediated NS2 trans-complementation, confirming that baculovirus expression of HCV proteins did not unduly affect particle production.Furthermore, this suggests that separation of structural protein expression from replicating HCV RNAs that are destined to be packaged alleviates an early stage requirement for NS2 during particle formation.This highlights our current lack of understanding of how NS2 mediates assembly, yet comparison of full-length and bipartite systems may provide further insight into this process.

View Article: PubMed Central - PubMed

Affiliation: Leeds Institute of Cancer & Pathology (LICAP), and Leeds CRUK Clinical Centre, Faculty of Medicine and Health, St James's University Hospital, University of Leeds, Beckett St., Leeds, West Yorkshire LS9 7TF, UK.

Show MeSH
Related in: MedlinePlus