Association of Potato Virus Y Gene Products with Chloroplasts in Tobacco

Gunasinghe, U. B.; Berger, P. H.

doi:10.1094/mpmi-4-452

Cited by 37 publications

(34 citation statements)

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“…Third, the fulllength negative-strand viral RNA was shown to be associated with the chloroplast-enriched fraction from TEV-infected tobacco (13). Finally, the viral genomic RNA was found in chloroplasts of tobacco plants infected with PVY (14). Collectively, these findings indicate that plant potyviruses initiate viral genome translation on the ER and form the 6K vesicles at ERES that traffic to chloroplasts for virus replication (Fig.…”

Section: Discussionmentioning

confidence: 76%

Sequential Recruitment of the Endoplasmic Reticulum and Chloroplasts for Plant Potyvirus Replication

Wèi

Huang

McNeil

et al. 2010

J Virol

200

225

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The replication of positive-strand RNA viruses occurs in cytoplasmic membrane-bound virus replication complexes (VRCs). Depending on the virus, distinct cellular organelles such as the endoplasmic reticulum (ER), chloroplast, mitochondrion, endosome, and peroxisome are recruited for the formation of VRC-associated membranous structures. Previously, the 6,000-molecular-weight protein (6K) of plant potyviruses was shown to be an integral membrane protein that induces the formation of 6K-containing membranous vesicles at endoplasmic reticulum (ER) exit sites for potyvirus genome replication. Here, we present evidence that the 6K-induced vesicles predominantly target chloroplasts, where they amalgamate and induce chloroplast membrane invaginations. The vesicular transport pathway and actomyosin motility system are involved in the trafficking of the 6K vesicles from the ER to chloroplasts. Viral RNA, double-stranded RNA, and viral replicase components are concentrated at the 6K vesicles that associate with chloroplasts in infected cells, suggesting that these chloroplast-bound 6K vesicles are the site for potyvirus replication. Taken together, these results suggest that plant potyviruses sequentially recruit the ER and chloroplasts for their genome replication.The replication of eukaryotic positive-strand RNA viruses in infected cells is closely associated with unique virus-induced intracellular membranous vesicles (22). These membranous vesicles have been proposed to provide a scaffold for anchoring the virus replication complex (VRC), confine the process of RNA replication to a specific safeguarded cytoplasmic location, and prevent the activation of certain host defense mechanisms that can be triggered by double-stranded RNA (dsRNA) intermediates during virus replication (33, 47). Depending on the type of virus, the virus-induced membranous vesicles are derived from various intracellular organelles in the host. Many plant and animal viruses remodel and utilize the endoplasmic reticulum (ER) in VRCs (1,6,17,33,34,36,38,39,46). Other cellular organelles such as endosomes, lysomes, chloroplasts, peroxisomes, and mitochondria have also been suggest to be the replication site for togaviruses, tymoviruses, and tombusviruses, respectively (25,27,31). Given that the ER appears to be the site where the host cell translation machinery is hijacked for the biosynthesis of the first set of viral proteins, the subcellular location of virus replication (either in the vicinity of the ER or elsewhere) and the mechanism of transport to locations other than the ER are poorly understood.Plant potyviruses, accounting for ϳ30% of known plant viruses including many agriculturally important viruses, e.g., Turnip mosaic virus (TuMV), Maize dwarf mosaic virus (MDMV), Tobacco etch virus (TEV), and Potato virus Y (PVY), are related to picornaviruses and picorna-like viruses (20,21,43). The potyviral genome is a single-stranded positive-sense RNA of about 10 kb in length and encodes at least 11 mature viral proteins (8, 43). Of these 11 protei...

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Section: Discussionmentioning

confidence: 76%

Sequential Recruitment of the Endoplasmic Reticulum and Chloroplasts for Plant Potyvirus Replication

Wèi

Huang

McNeil

et al. 2010

J Virol

200

225

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“…In addition, Turnip mosaic virus HC-Pro and Potato virus Y (PVY) HC-Pro could form aggregates in the cytoplasm (Zheng et al, 2011;del Toro et al, 2014), and an amino acid change of Papaya ringspot virus HC-Pro also led to the formation of the aggregates (Sahana et al, 2014). Interestingly, PVY HC-Pro also could be present in the chloroplasts of PVY-infected plants (Gunasinghe and Berger, 1991). This discrepancy of HC-Pro distribution may be due to different characteristics of specific viruses.…”

Section: Discussionmentioning

confidence: 99%

“…Approximately 9 g of maize seedling leaves was used for chloroplast isolation. The reconstitution experiments were performed using chloroplasts isolated from healthy maize plants mixed with the supernatant of a chloroplast preparation from the same amount of SCMV-infected maize tissues as described previously (Gunasinghe and Berger, 1991) to rule out the contamination by cytosol proteins. For chloroplasts from reconstitution experiments and SCMV-infected maize leaves, thermolysin (Promega) treatment (Cline et al, 1984) was conducted as described previously to digest the proteins attached to the surface of chloroplasts.…”

Section: Maize Chloroplast Isolation and Protein Extractionmentioning

confidence: 99%

A Violaxanthin Deepoxidase Interacts with a Viral Suppressor of RNA Silencing to Inhibit Virus Amplification

et al. 2017

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RNA silencing plays a critical role against viral infection. To counteract this antiviral silencing, viruses have evolved various RNA silencing suppressors. Meanwhile, plants have evolved counter-counter defense strategies against RNA silencing suppression (RSS). In this study, the violaxanthin deepoxidase protein of maize (Zea mays), ZmVDE, was shown to interact specifically with the helper component-proteinase (HC-Pro; a viral RNA silencing suppressor) of Sugarcane mosaic virus (SCMV) via its mature protein region by yeast two-hybrid assay, which was confirmed by coimmunoprecipitation in Nicotiana benthamiana cells. It was demonstrated that amino acids 101 to 460 in HC-Pro and the amino acid glutamine-292 in ZmVDE mature protein were essential for this interaction. The mRNA levels of ZmVDE were down-regulated 75% to 65% at an early stage of SCMV infection. Interestingly, ZmVDE, which normally localized in the chloroplasts and cytoplasm, could relocalize to HC-Pro-containing aggregate bodies in the presence of HC-Pro alone or SCMV infection. In addition, ZmVDE could attenuate the RSS activity of HC-Pro in a specific protein interaction-dependent manner. Subsequently, transient silencing of the ZmVDE gene facilitated SCMV RNA and coat protein accumulation. Taken together, our results suggest that ZmVDE interacts with SCMV HC-Pro and attenuates its RSS activity, contributing to decreased SCMV accumulation. This study demonstrates that a host factor can be involved in secondary defense responses against viral infection in monocot plants.

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“…These symptoms might be explained by the presence of the PVY-CP in chloroplasts of infected leaves (Gunasinghe and Berger, 1991), since transgenic plants expressing the PVY-CP in their chloroplasts loose their green color and grow very badly (Naderi and Berger, 1997). These plants have the same phenotype as the mutant N. tabacum that expresses low RubisCO levels associated with chlorophyll loss (Dulieu, 1974;Nguyen-Quoc et al, 1989).…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, an amino acid triplet, Asp-Ala-Gly (DAG), near the surface-exposed N-terminus of the CP (Shukla and Ward, 1989) is highly conserved among aphidtransmissible potyviruses and involved in the interaction with the helper component (HC) during aphid transmission (Atreya et al, 1995;Blanc et al, 1997). The PVY symptoms might be explained by the presence of CP, HC and RNA of this virus in chloroplasts of infected leaves (Gunasinghe and Berger, 1991), which might interfere with the chloroplastic function including RubisCO photosynthetic activity (Miziorko and Lorimer, 1983;Suzuki, 1987;Rival et al, 1996). This coincides with the fact that transgenic plants expressing PVY-CP in their chloroplasts lose their green color and grow poorly (Naderi and Berger, 1997).…”

Section: Introductionmentioning

confidence: 99%

Interaction between tobacco Ribulose-l,5-biphosphate Carboxylase/Oxygenase large subunit (RubisCO-LSU) and the PVY Coat Protein (PVY-CP)

et al. 2005

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A 54 kDa band (P54) was continually detected with the 30 kDa viral capsid protein (CP) on the SDS-PAGE migration profile of purified potato virus Y (PVY). P54 was observed following the use of two different procedures for the purification of the PVY from infected tobacco. It was a constitutively expressed tobacco protein. The analysis of the PVY preparation showed that P54 has aggregation properties and precipitates, thus pulling down the virus. We used an enzyme-linked immunosorbent assay (ELISA) to study the relationship between P54 and the PVY particles. We performed an inhibition test with monoclonal antibodies (mAb) directed against the PVY-CP, to show that these two components interact. This result was confirmed by western blot. The internal sequence of five major peptides, obtained by C-lysine endoprotease digestion of the P54 followed by HPLC separation, showed 100% homology with the large subunit of the ribulose-1,5-biphosphate carboxylase/oxygenase (RubisCO-LSU) of tobacco. MAb directed against RubisCO-LSU were produced and used to reveal the RubisCO-LSU/PVY complex in infected tobacco extracts. A phage library displaying random heptapeptides was used to isolate several peptides that specifically bound to the native form of the PVY. The sequences of thirty-three phage-displayed peptides, which bound specifically to this virus, present further discontinuous sequence homologies with the RubisCO-LSU. Five peptides (p1 to p5) corresponding to the RubisCO-LSU homologous regions were used for a bacterial two-hybrid system to confirm in vivo direct interactions between the selected RubisCO-LSU regions and the PVY-CP. We propose that the PVY-CP may be involved in the production of mosaics and yellowing symptoms in tobacco through its interaction with RubisCO-LSU. Abbreviations: AC

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Association of Potato Virus Y Gene Products with Chloroplasts in Tobacco

Cited by 37 publications

References 0 publications

Sequential Recruitment of the Endoplasmic Reticulum and Chloroplasts for Plant Potyvirus Replication

Sequential Recruitment of the Endoplasmic Reticulum and Chloroplasts for Plant Potyvirus Replication

A Violaxanthin Deepoxidase Interacts with a Viral Suppressor of RNA Silencing to Inhibit Virus Amplification

Interaction between tobacco Ribulose-l,5-biphosphate Carboxylase/Oxygenase large subunit (RubisCO-LSU) and the PVY Coat Protein (PVY-CP)

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