Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection

Nicholas S. Eyre, Guillaume N. Fiches, Amanda L. Aloia, Karla J. Helbig, Erin M. McCartney, Christopher S.P. McErlean, Kui Li, Anupriya Aggarwal, Stuart G. Turville, Michael R. Beard

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Hepatitis C virus (HCV) NS5A is essential for viral genome replication within cytoplasmic replication complexes and virus assembly at the lipid droplet (LD) surface, although its definitive functions are poorly understood. We developed approaches to investigate NS5A dynamics during a productive infection. We report here that NS5A motility and efficient HCV RNA replication require the microtubule network and the cytoplasmic motor dynein and demonstrate that both motile and relatively static NS5A-positive foci are enriched with host factors VAP-A and Rab5A. Pulse-chase imaging revealed that newly synthesized NS5A foci are small and distinct from aged foci, while further studies using a unique dual fluorescently tagged infectious HCV chimera showed a relatively stable association of NS5A foci with core-capped LDs. These results reveal new details about the dynamics and maturation of NS5A and the nature of potential sites of convergence of HCV replication and assembly pathways.

Original languageEnglish (US)
Pages (from-to)3636-3652
Number of pages17
JournalJournal of Virology
Volume88
Issue number7
DOIs
StatePublished - Jan 1 2014

Fingerprint

Hepatitis C virus
Hepacivirus
image analysis
Virus Assembly
Virus Replication
Infection
infection
Proteins
Hepatovirus
proteins
Cytoplasmic Dyneins
hepatitis A
chimerism
Viral Genome
virus replication
Microtubules
microtubules
droplets
RNA
genome

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Eyre, N. S., Fiches, G. N., Aloia, A. L., Helbig, K. J., McCartney, E. M., McErlean, C. S. P., ... Beard, M. R. (2014). Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection. Journal of Virology, 88(7), 3636-3652. https://doi.org/10.1128/JVI.02490-13

Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection. / Eyre, Nicholas S.; Fiches, Guillaume N.; Aloia, Amanda L.; Helbig, Karla J.; McCartney, Erin M.; McErlean, Christopher S.P.; Li, Kui; Aggarwal, Anupriya; Turville, Stuart G.; Beard, Michael R.

In: Journal of Virology, Vol. 88, No. 7, 01.01.2014, p. 3636-3652.

Research output: Contribution to journalArticle

Eyre, NS, Fiches, GN, Aloia, AL, Helbig, KJ, McCartney, EM, McErlean, CSP, Li, K, Aggarwal, A, Turville, SG & Beard, MR 2014, 'Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection', Journal of Virology, vol. 88, no. 7, pp. 3636-3652. https://doi.org/10.1128/JVI.02490-13
Eyre NS, Fiches GN, Aloia AL, Helbig KJ, McCartney EM, McErlean CSP et al. Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection. Journal of Virology. 2014 Jan 1;88(7):3636-3652. https://doi.org/10.1128/JVI.02490-13
Eyre, Nicholas S. ; Fiches, Guillaume N. ; Aloia, Amanda L. ; Helbig, Karla J. ; McCartney, Erin M. ; McErlean, Christopher S.P. ; Li, Kui ; Aggarwal, Anupriya ; Turville, Stuart G. ; Beard, Michael R. / Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection. In: Journal of Virology. 2014 ; Vol. 88, No. 7. pp. 3636-3652.
@article{94cb438ec18f4fb6b86469230d8499ac,
title = "Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection",
abstract = "Hepatitis C virus (HCV) NS5A is essential for viral genome replication within cytoplasmic replication complexes and virus assembly at the lipid droplet (LD) surface, although its definitive functions are poorly understood. We developed approaches to investigate NS5A dynamics during a productive infection. We report here that NS5A motility and efficient HCV RNA replication require the microtubule network and the cytoplasmic motor dynein and demonstrate that both motile and relatively static NS5A-positive foci are enriched with host factors VAP-A and Rab5A. Pulse-chase imaging revealed that newly synthesized NS5A foci are small and distinct from aged foci, while further studies using a unique dual fluorescently tagged infectious HCV chimera showed a relatively stable association of NS5A foci with core-capped LDs. These results reveal new details about the dynamics and maturation of NS5A and the nature of potential sites of convergence of HCV replication and assembly pathways.",
author = "Eyre, {Nicholas S.} and Fiches, {Guillaume N.} and Aloia, {Amanda L.} and Helbig, {Karla J.} and McCartney, {Erin M.} and McErlean, {Christopher S.P.} and Kui Li and Anupriya Aggarwal and Turville, {Stuart G.} and Beard, {Michael R.}",
year = "2014",
month = "1",
day = "1",
doi = "10.1128/JVI.02490-13",
language = "English (US)",
volume = "88",
pages = "3636--3652",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "7",

}

TY - JOUR

T1 - Dynamic imaging of the hepatitis C virus NS5A protein during a productive infection

AU - Eyre, Nicholas S.

AU - Fiches, Guillaume N.

AU - Aloia, Amanda L.

AU - Helbig, Karla J.

AU - McCartney, Erin M.

AU - McErlean, Christopher S.P.

AU - Li, Kui

AU - Aggarwal, Anupriya

AU - Turville, Stuart G.

AU - Beard, Michael R.

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Hepatitis C virus (HCV) NS5A is essential for viral genome replication within cytoplasmic replication complexes and virus assembly at the lipid droplet (LD) surface, although its definitive functions are poorly understood. We developed approaches to investigate NS5A dynamics during a productive infection. We report here that NS5A motility and efficient HCV RNA replication require the microtubule network and the cytoplasmic motor dynein and demonstrate that both motile and relatively static NS5A-positive foci are enriched with host factors VAP-A and Rab5A. Pulse-chase imaging revealed that newly synthesized NS5A foci are small and distinct from aged foci, while further studies using a unique dual fluorescently tagged infectious HCV chimera showed a relatively stable association of NS5A foci with core-capped LDs. These results reveal new details about the dynamics and maturation of NS5A and the nature of potential sites of convergence of HCV replication and assembly pathways.

AB - Hepatitis C virus (HCV) NS5A is essential for viral genome replication within cytoplasmic replication complexes and virus assembly at the lipid droplet (LD) surface, although its definitive functions are poorly understood. We developed approaches to investigate NS5A dynamics during a productive infection. We report here that NS5A motility and efficient HCV RNA replication require the microtubule network and the cytoplasmic motor dynein and demonstrate that both motile and relatively static NS5A-positive foci are enriched with host factors VAP-A and Rab5A. Pulse-chase imaging revealed that newly synthesized NS5A foci are small and distinct from aged foci, while further studies using a unique dual fluorescently tagged infectious HCV chimera showed a relatively stable association of NS5A foci with core-capped LDs. These results reveal new details about the dynamics and maturation of NS5A and the nature of potential sites of convergence of HCV replication and assembly pathways.

UR - http://www.scopus.com/inward/record.url?scp=84895476594&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84895476594&partnerID=8YFLogxK

U2 - 10.1128/JVI.02490-13

DO - 10.1128/JVI.02490-13

M3 - Article

C2 - 24429364

AN - SCOPUS:84895476594

VL - 88

SP - 3636

EP - 3652

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 7

ER -