Abstract
Superinfection exclusion (SIE) is an antagonistic interaction between identical or closely related viruses in host cells. Previous studies by us and others led to the hypothesis that SIE was elicited by one or more proteins encoded in the genomes of primary viruses. Here, we tested this hypothesis using Turnip mosaic virus (TuMV), a member of the genus Potyvirus of the family Potyviridae, with significant economic consequences. To this end, individual TuMV-encoded proteins were transiently expressed in the cells of Nicotiana benthamiana leaves, followed by challenging them with a modified TuMV expressing the green fluorescent protein (TuMV-GFP). Three days after TuMV-GFP delivery, these cells were examined for the replication-dependent expression of GFP. Cells expressing TuMV P1, HC-Pro, 6K1, CI, 6K2, NIa-VPg, NIb, or CP proteins permitted an efficient expression of GFP, suggesting that these proteins failed to block the replication of a superinfecting TuMV-GFP. By contrast, N. benthamiana cells expressing TuMV P3 or NIa-Pro did not express visible GFP fluorescence, suggesting that both of them could elicit potent SIE against TuMV-GFP. The SIE elicitor activity of P3 and NIa-Pro was further confirmed by their heterologous expression from a different potyvirus, potato virus A (PVA). Plants systemically infected with PVA variants expressing TuMV P3 or NIa-Pro blocked subsequent infection by TuMV-GFP. A +1-frameshift mutation in P3 and NIa-Pro cistrons facilitated superinfection by TuMV-GFP, suggesting that the P3 and NIa-Pro proteins, but not the RNA, are involved in SIE activity. Additionally, deletion mutagenesis identified P3 amino acids 3 to 200 of 352 and NIa-Pro amino acids 3 to 40 and 181 to 242 of 242 as essential for SIE elicitation. Collectively, our study demonstrates that TuMV encodes two spatially separated proteins that act independently to exert SIE on superinfecting TuMV. These results lay the foundation for further mechanistic interrogations of SIE in this virus.
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Claims 9
Claim · model-written summary
Cells expressing eight other TuMV proteins permitted efficient GFP expression.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianat0 - Interactor
- TuMV
Turnip mosaic viruse0pathogen - Gene
- TuMV P1, HC-Pro, 6K1, CI, 6K2, NIa-VPg, NIb, or CP proteinse0
- Outcome
- efficient expression of GFPe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Cells expressing TuMV P1, HC-Pro, 6K1, CI, 6K2, NIa-VPg, NIb, or CP proteins permitted an efficient expression of GFP”
t0“Nicotiana benthamiana”
Claim · model-written summary
Cells expressing TuMV P3 showed no visible GFP fluorescence after TuMV-GFP challenge.
- Taxon
- N. benthamianaNicotiana benthamianae0
- Interactor
- TuMVTurnip mosaic viruse0pathogen
- Gene
- TuMV P3e0
- Outcome
- did not express visible GFP fluorescencee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“N. benthamiana cells expressing TuMV P3 or NIa-Pro did not express visible GFP fluorescence”
Claim · model-written summary
Cells expressing NIa-Pro showed no visible GFP fluorescence after TuMV-GFP challenge.
- Taxon
- N. benthamianaNicotiana benthamianae0
- Interactor
- TuMVTurnip mosaic viruse0pathogen
- Gene
- NIa-Proe0
- Outcome
- did not express visible GFP fluorescencee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“N. benthamiana cells expressing TuMV P3 or NIa-Pro did not express visible GFP fluorescence”
Claim · model-written summary
PVA variants expressing TuMV P3 blocked subsequent TuMV-GFP infection.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- PVA variants expressing TuMV P3e0
- Interactor
- TuMV-GFPe0pathogen
- Second interactor
- PVAPotato virus Ae0pathogen
- Gene
- TuMV P3e0
- Outcome
- infection by TuMV-GFPe0
- Direction
- blockede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Plants systemically infected with PVA variants expressing TuMV P3 or NIa-Pro blocked subsequent infection by TuMV-GFP”
t0“Nicotiana benthamiana”
Claim · model-written summary
PVA variants expressing TuMV NIa-Pro blocked subsequent TuMV-GFP infection.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- PVA variants expressing TuMV P3 or NIa-Proe0
- Interactor
- TuMV-GFPe0pathogen
- Second interactor
- PVAPotato virus Ae0pathogen
- Gene
- NIa-Proe0
- Outcome
- infection by TuMV-GFPe0
- Direction
- blockede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Plants systemically infected with PVA variants expressing TuMV P3 or NIa-Pro blocked subsequent infection by TuMV-GFP”
t0“Nicotiana benthamiana”
Claim · model-written summary
A +1 frameshift in P3 facilitated TuMV-GFP superinfection.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- +1-frameshift mutation in P3e0
- Interactor
- TuMV-GFPe0pathogen
- Gene
- P3e0
- Outcome
- superinfection by TuMV-GFPe0
- Direction
- facilitatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A +1-frameshift mutation in P3 and NIa-Pro cistrons facilitated superinfection by TuMV-GFP”
t0“Nicotiana benthamiana”
Claim · model-written summary
A +1 frameshift in NIa-Pro facilitated TuMV-GFP superinfection.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- +1-frameshift mutation in P3 and NIa-Pro cistronse0
- Interactor
- TuMV-GFPe0pathogen
- Gene
- NIa-Proe0
- Outcome
- superinfection by TuMV-GFPe0
- Direction
- facilitatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A +1-frameshift mutation in P3 and NIa-Pro cistrons facilitated superinfection by TuMV-GFP”
t0“Nicotiana benthamiana”
Claim · model-written summary
P3 amino acids 3 to 200 of 352 were essential for SIE.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Gene
- P3e0
- Outcome
- essential for SIE elicitatione0
- Qualifier
- amino acids 3 to 200 of 352e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“P3 amino acids 3 to 200 of 352 and NIa-Pro amino acids 3 to 40 and 181 to 242 of 242 as essential for SIE elicitation”
t0“Nicotiana benthamiana”
Claim · model-written summary
NIa-Pro amino acids 3 to 40 and 181 to 242 of 242 were essential for SIE.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Gene
- NIa-Proe0
- Outcome
- essential for SIE elicitatione0
- Qualifier
- amino acids 3 to 40 and 181 to 242 of 242e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“NIa-Pro amino acids 3 to 40 and 181 to 242 of 242 as essential for SIE elicitation”
t0“Nicotiana benthamiana”
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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.