Abstract
The formation of virus movement protein (MP)-containing punctate structures on the cortical endoplasmic reticulum is required for efficient intercellular movement of Red clover necrotic mosaic virus (RCNMV), a bipartite positive-strand RNA plant virus. We found that these cortical punctate structures constitute a viral replication complex (VRC) in addition to the previously reported aggregate structures that formed adjacent to the nucleus. We identified host proteins that interacted with RCNMV MP in virus-infected Nicotiana benthamiana leaves using a tandem affinity purification method followed by mass spectrometry. One of these host proteins was glyceraldehyde 3-phosphate dehydrogenase-A (NbGAPDH-A), which is a component of the Calvin-Benson cycle in chloroplasts. Virus-induced gene silencing of NbGAPDH-A reduced RCNMV multiplication in the inoculated leaves, but not in the single cells, thereby suggesting that GAPDH-A plays a positive role in cell-to-cell movement of RCNMV. The fusion protein of NbGAPDH-A and green fluorescent protein localized exclusively to the chloroplasts. In the presence of RCNMV RNA1, however, the protein localized to the cortical VRC as well as the chloroplasts. Bimolecular fluorescence complementation assay and GST pulldown assay confirmed in vivo and in vitro interactions, respectively, between the MP and NbGAPDH-A. Furthermore, gene silencing of NbGAPDH-A inhibited MP localization to the cortical VRC. We discuss the possible roles of NbGAPDH-A in the RCNMV movement process.
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Claims 9
Claim · model-written summary
Cortical punctate structures containing the movement protein constitute a viral replication complex of RCNMV, in addition to aggregate structures adjacent to the nucleus.
- Interactor
- Red clover necrotic mosaic virusRed clover necrotic mosaic viruse1pathogen
- Outcome
- viral replication complexe0
Not reportedTaxon
Verbatim
e0“these cortical punctate structures constitute a viral replication complex (VRC)”
e1“Red clover necrotic mosaic virus”
Claim · model-written summary
NbGAPDH-A was identified as a host protein interacting with the RCNMV movement protein in infected Nicotiana benthamiana leaves.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae0 - Organ / stage
- leavese0
- Interactor
- RCNMVRed clover necrotic mosaic viruse0pathogen
- Gene
- NbGAPDH-Ae1
- Outcome
- host proteins that interacted with RCNMV MPe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We identified host proteins that interacted with RCNMV MP in virus-infected Nicotiana benthamiana leaves”
e1“glyceraldehyde 3-phosphate dehydrogenase-A (NbGAPDH-A)”
Claim · model-written summary
Virus-induced gene silencing of NbGAPDH-A reduced RCNMV multiplication in inoculated leaves.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- Virus-induced gene silencing of NbGAPDH-Ae0
- Interactor
- RCNMVRed clover necrotic mosaic viruse0pathogen
- Gene
- NbGAPDH-Ae0
- Outcome
- RCNMV multiplication in the inoculated leavese0
- Direction
- reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Virus-induced gene silencing of NbGAPDH-A reduced RCNMV multiplication in the inoculated leaves”
e1“virus-infected Nicotiana benthamiana leaves”
Claim · model-written summary
Silencing of NbGAPDH-A did not reduce RCNMV multiplication in single cells.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae2
- Treatment
- Virus-induced gene silencing of NbGAPDH-Ae1
- Interactor
- RCNMVRed clover necrotic mosaic viruse0pathogen
- Outcome
- RCNMV multiplicatione0
- Direction
- not in the single cellse0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“reduced RCNMV multiplication in the inoculated leaves, but not in the single cells”
e1“Virus-induced gene silencing of NbGAPDH-A”
e2“virus-infected Nicotiana benthamiana leaves”
Claim · model-written summary
The authors suggest that GAPDH-A plays a positive role in cell-to-cell movement of RCNMV.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Interactor
- RCNMVRed clover necrotic mosaic viruse0pathogen
- Gene
- GAPDH-Ae0
- Outcome
- cell-to-cell movement of RCNMVe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“thereby suggesting that GAPDH-A plays a positive role in cell-to-cell movement of RCNMV”
e1“virus-infected Nicotiana benthamiana leaves”
Claim · model-written summary
The NbGAPDH-A-GFP fusion protein localized exclusively to chloroplasts.
Not reportedTaxonDirection
Verbatim
e0“The fusion protein of NbGAPDH-A and green fluorescent protein localized exclusively to the chloroplasts”
Claim · model-written summary
In the presence of RCNMV RNA1, the NbGAPDH-A fusion protein localized to the cortical VRC as well as the chloroplasts.
- Outcome
- localized to the cortical VRC as well as the chloroplastse0
- Qualifier
- In the presence of RCNMV RNA1e0
Not reportedTaxonDirection
Verbatim
e0“In the presence of RCNMV RNA1, however, the protein localized to the cortical VRC as well as the chloroplasts”
Claim · model-written summary
Bimolecular fluorescence complementation and GST pulldown assays confirmed in vivo and in vitro interactions, respectively, between the RCNMV movement protein and NbGAPDH-A.
Not reportedTaxonDirection
Verbatim
e0“Bimolecular fluorescence complementation assay and GST pulldown assay confirmed in vivo and in vitro interactions, respectively, between the MP and NbGAPDH-A”
Claim · model-written summary
Gene silencing of NbGAPDH-A inhibited localization of the movement protein to the cortical VRC.
- Treatment
- gene silencing of NbGAPDH-Ae0
- Gene
- NbGAPDH-Ae0
- Outcome
- MP localization to the cortical VRCe0
- Direction
- inhibitede0
Not reportedTaxonComparator
Verbatim
e0“gene silencing of NbGAPDH-A inhibited MP localization to the cortical VRC”
The same questions in other papers
loss of function · quality: protein · Nicotiana benthamianaPositive and negative results
3 papers in the same direction as this paper · 10 in the opposite direction
- negative
- negative
- negative
10 papers more in this group
See all 14 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.