Subcellular localization and live-cell imaging of the Helicoverpa armigera stunt virus replicase in mammalian and Spodoptera frugiperda cells

Short, James R.; Knox, Caroline; Dorrington, Rosemary A.

doi:10.1099/vir.0.020156-0

Cited by 8 publications

(16 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Three distinct distributions of EGFP fluorescence were observed in cells expressing these mutants ( Fig. 2a): (i) EGFP fluorescence distributed throughout the cytoplasm that appeared to concentrate within the nucleus ('Cytosolic+Nuc'); (ii) diffuse cytosolic fluorescence, excluded from the nucleus ('Cytosolic -Nuc'); and (iii) the 'Punctate' localized distribution as previously reported for REP 1-1704 -EGFP (Short et al, 2010). The proportion of cells displaying each dominant distribution, expressed as a percentage of the total number of cells counted, was also determined (Fig.…”

Section: Resultssupporting

confidence: 71%

Membrane targeting of an alpha-like tetravirus replicase is directed by a region within the RNA-dependent RNA polymerase domain

Short

Dorrington

2012

Journal of General Virology

View full text Add to dashboard Cite

The members of the family Tetraviridae are small positive-sense insect RNA viruses that exhibit stringent host specificity and a high degree of tissue tropism, suggesting that complex virus-host interactions are likely to occur during infection and viral replication. The alpha-like replicase of Helicoverpa armigera stunt virus (HaSV) (genus Omegatetravirus) has been proposed to associate with membranes of the endocytic pathway, which is similar to Semliki Forest virus, Sindbis virus and rubella virus. Here, we have used replicase-EGFP fusion proteins and recombinant baculovirus expression to demonstrate that the HaSV replicase associates strongly with cellular membranes, including detergent-resistant membranes, and that this association is maintained through a novel membrane targeting domain within the C-terminal region of the RNA-dependent RNA polymerase domain. We show a similar subcellular localization and strong association with detergent-resistant membranes for the carmo-like replicase of another tetravirus, Providence virus, in replicating cells, suggesting a common site of replication for these two tetraviruses.

show abstract

Section: Resultssupporting

confidence: 71%

Membrane targeting of an alpha-like tetravirus replicase is directed by a region within the RNA-dependent RNA polymerase domain

Short

Dorrington

2012

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…These biochemical features indicate that DpTV Hel should function in a limited environment in cells. Indeed, RNA replication of ( þ)RNA viruses requires viral RNA replicase proteins to associate with intracellular membranes and induce membrane invagination to form small spherules (Ahlquist, 2006;Short and Dorrington, 2012;Short et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…For alphatetraviruses, they encode a large viral RNA replicase protein (DpTV: $ 180 kDa, HaSV: $187 kDa, NbV: $ 215 kDa). On the basis of the amino acid sequences, some alphatetraviral replicases, including NbV and all omegatetraviruses, were predicted to contain three putative domains indicative of methyltransferase (MT), RNA helicase (Hel), and RNA-dependent RNA polymerase (RdRp) (Gordon et al, 1995Hanzlik and Gordon, 1997;Pringle et al, 1999;Short et al, 2010;Yi et al, 2005;Zeddam et al, 2010). Despite the importance of helicases in (þ)RNA viruses, helicase activity in alphatetraviruses has not been formally determined, and consequently, the molecular mechanisms of helicasemediated unwinding of alphatetraviral dsRNA have not yet been studied, which hinders our understanding of this family of viruses.…”

Section: Introductionmentioning

confidence: 99%

Identification and characterization of RNA duplex unwinding and ATPase activities of an alphatetravirus superfamily 1 helicase

et al. 2012

View full text Add to dashboard Cite

Dendrolimus punctatus tetravirus (DpTV) belongs to the genus omegatetravirus of the Alphatetraviridae family. Sequence analysis predicts that DpTV replicase contains a putative helicase domain (Hel). However, the helicase activity in alphatetraviruses has never been formally determined. In this study, we determined that DpTV Hel is a functional RNA helicase belonging to superfamily-1 helicase with 5'-3' dsRNA unwinding directionality. Further characterization determined the length requirement of the 5' single-stranded tail on the RNA template and the optimal reaction conditions for the unwinding activity of DpTV Hel. Moreover, DpTV Hel also contains NTPase activity. The ATPase activity of DpTV Hel could be significantly stimulated by dsRNA, and dsRNA could partially rescue the ATPase activity abolishment caused by mutations. Our study is the first to identify an alphatetravirus RNA helicase and further characterize its dsRNA unwinding and NTPase activities in detail and should foster our understanding of DpTV and other alphatetraviruses.

show abstract

“…Dextrans are hydrophilic polysaccharides that are characterized by a range of molecular weights and low toxicity (Nouvel et al ., ). Fluorescently‐labeled dextrans (Molecular Probes, Carlsbad, CA, USA) are routinely used to analyze macropinocytosis and fluid‐phase endocytosis in a variety of cell and tissue types (Masedunskas & Weigert, ; Petrescu et al ., ; Short et al ., ).…”

Section: Introductionmentioning

confidence: 97%

Localization of Banana bunchy top virus and cellular compartments in gut and salivary gland tissues of the aphid vector Pentalonia nigronervosa

2015

View full text Add to dashboard Cite

Banana bunchy top virus (BBTV) (Nanoviridae: Babuvirus) is transmitted by aphids of the genus Pentalonia in a circulative manner. The cellular mechanisms by which BBTV translocates from the anterior midgut to the salivary gland epithelial tissues are not understood. Here, we used multiple fluorescent markers to study the distribution and the cellular localization of early and late endosomes, macropinosomes, lysosomes, microtubules, actin filaments, and lipid raft subdomains in the gut and principal salivary glands of Pentalonia nigronervosa. We applied colabeling assays, to colocalize BBTV viral particles with these cellular compartments and structures. Our results suggest that multiple potential cellular processes, including clathrin- and caveolae-mediated endocytosis and lipid rafts, may not be involved in BBTV internalization.

show abstract

Subcellular localization and live-cell imaging of the Helicoverpa armigera stunt virus replicase in mammalian and Spodoptera frugiperda cells

Cited by 8 publications

References 39 publications

Membrane targeting of an alpha-like tetravirus replicase is directed by a region within the RNA-dependent RNA polymerase domain

Membrane targeting of an alpha-like tetravirus replicase is directed by a region within the RNA-dependent RNA polymerase domain

Identification and characterization of RNA duplex unwinding and ATPase activities of an alphatetravirus superfamily 1 helicase

Localization of Banana bunchy top virus and cellular compartments in gut and salivary gland tissues of the aphid vector Pentalonia nigronervosa

Contact Info

Product

Resources

About