A viral protein promotes host SAMS1 activity and ethylene production for the benefit of virus infection
- Peking University, China
- The First Affiliated Hospital of Zhejiang Chinese Medical University, China
- Fujian Agriculture and Forestry University, China
- Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, China
Figures
Figure 1 with 1 supplement
Pns11 overexpression lines are more susceptible to RDV infection than WT plants.
(A) Symptoms of the mock-inoculated WT or RDV-infected WT (non-transgenic) plants and S11 OX transgenic plants; images were taken at 4 wpi. Scale bars = 10 cm (upper panel) and 5 cm (lower panel). (B…
Figure 1—figure supplement 1
Phenotype and identification of S11-overexpressing lines.
(A) Expression levels of S11 driven by the Ubiquitin promoter in transgenic rice lines (#3, #5, and #11). Actin was used as an internal control. (B) Western blot analysis of Pns11 protein expression …
Figure 2 with 7 supplements
Pns11 interacts with OsSAMS1 and enhances its activity.
(A) Yeast two-hybrid assay for the interaction of Pns11 with OsSAMS1. The bait vector contained full-length Pns11; the prey vector contained OsSAMS1, OsSAMS2, or OsSAMS3. Yeast strains were cultured …
Figure 2—figure supplement 1
RDV Pns11 specifically interacts with OsSAMS1 and does not affect OsSAMS1 expression.
(A) Pns11 and OsSAMS1 specifically interact with each other in a yeast two-hybrid assay. (B) Co-IP assay for the interaction of Pns11 with OsSAMS2. Pns11 and OsSAMS2 proteins were transiently …
Figure 2—figure supplement 2
Pns11 and OsSAMS1 are co-localized in both nucleus and cytoplasm.
Bimolecular fluorescence complementation (BiFC) interaction. Constructs expressing Pns11 and OsSAMS1 fused to the N- or C-terminal portions of yellow fluorescent protein (YFP) were delivered to N. …
Figure 2—figure supplement 3
Neither GFP nor P9 affects OsSAMS1 activity in vitro.
(A) The upper graph represents the coomassie brilliant blue staining of varying amounts of OsSAMS1 and GFP, respectively. The middle graph is an autoradiograph of a representative chromatogram …
Figure 2—figure supplement 4
Pns11 does not affect OsSAMS2 activity in vitro.
(A) Coomassie brilliant blue staining of varying amounts of OsSAMS2 and Pns11, respectively. (B) Autoradiograph of a representative chromatogram showing SAM generated by OsSAMS2 in reactions …
Figure 2—figure supplement 5
Pns11 does not affect the affinity of OsSAMS1 to the substrates L-Met or ATP.
(A) Pns11 does not affect dynamic association between OsSAMS1 and L-Met. Data were collected from microscale thermophoresis (MST) assays as described in the 'Materials and methods'. Experiments were …
Figure 2—figure supplement 6
Multiple alignments of SAM synthetase proteins.
Multiple alignments of SAM synthetase proteins. The amino acid secquence of OsSASM1 synthetase protein (Q0DKY4.1) were aligned with Oryza sativa SAM synthetase 2 (OsSAMS2, P93438.1), O. sativa SAM …
Figure 2—figure supplement 7
Gel-filtration analysis of Pns11 and OsSAMS1.
Gel filtration analysis of Pns11 and OsSAMS1. The high-molecular-weight fraction peak of Pns11 and OsSAMS1 is indicated. The two vertical black arrows indicate fractions in which the 669-kDa and …
Figure 3 with 2 supplements
SAM, ACC, and ethylene contents in OsSAMS1 and S11 transgenic lines.
(A) SAM contents in the S11-overexpression lines and WT plants (using 40-d-old seedlings). (B) ACC contents in the S11-overexpression lines and WT plants (using 40-d-old seedlings). (C) Ethylene …
Figure 3—figure supplement 1
Phenotype and identification of OsSAMS1 transgenic lines.
(A) Expression levels of OsSAMS1 in WT, OX and RNAi transgenic lines. (14-d-old seedlings). The average (± standard deviation) values from three biological replicates are shown. Significant …
Figure 3—figure supplement 2
SAM, ACC and ethylene contents in OsSAMS1 transgenic lines.
(A) SAM content in OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). (B) ACC content in OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). (C) Ethylene content in OsS…
Figure 4 with 1 supplement
Overexpression of OsSAMS1 enhances RDV infection whereas knockout of OsSAMS1 reduces RDV infection.
(A) Symptoms of the mock-inoculated WT or RDV-infected WT (non-transgenic) plants and OsSAMS1 OX/KO transgenic plants; images were taken at 4 wpi. Scale bars = 10 cm (upper panel) and 5 cm (lower …
Figure 4—figure supplement 1
Overexpression of OsSAMS1 enhances RDV infection while downregulation of OsSAMS1 reduces RDV infection.
(A) Symptoms of mock or RDV-infected WT plants as well as OsSAMS1 OX/RNAi transgenic plants, pictures were taken at 4 wpi. Scale bars, 10 cm (upper panel) and 5 cm (lower panel). (B) Detection of …
Figure 5 with 3 supplements
The ethylene-response mutant (osein2) shows increased tolerance of RDV infection whereas overexpression (OX) of OsEIN2 results in enhanced susceptibility.
(A) Symptoms of the mock-inoculated WT or RDV-infected WT (non-transgenic) plants, osein2, and OsEIN2-overexpression (OX) plants; images were taken at 4 wpi. Scale bars = 10 cm (upper panel) and 5 …
Figure 5—figure supplement 1
Phenotype and identification of OsSAMS1 OE/osein2 (J119) transgenic lines.
(A) Confirmation of osein2 and J119 lines by PCR-based analysis. (B) Western blot analysis of OsSAMS1 protein expression level in transgenic lines. Actin was probed and served as a loading control. …
Figure 5—figure supplement 2
Overexpressed OsSAMS1 in osein2 (J119) results in increased tolerance of RDV infection.
(A) Symptoms of the mock-inoculated WT or RDV-infected WT (non-transgenic), osein2 and J119 plants; images were taken at 4 wpi. Scale bars = 10 cm (upper panel) and 5 cm (lower panel). (B) Detection …
Figure 5—figure supplement 3
Detection of salicylic acid (SA)-, jasmonic acid (JA)- and ethylene-responsive genes.
(A) Expression of SA-, JA- and ethylene-responsive genes over time after RDV infection in NPB, OSEIN2 OX#2 and osein2 rice lines. (B) Expression of SA-, JA- and ethylene-responsive genes over time …
Figure 6 with 1 supplement
Ethylene is induced by RDV infection, which enhances the susceptibility of rice to RDV infection.
(A) In vivo pull-down in WT and Ossams1 KO lines, with or without RDV infection (40-d-old seedlings, 4 wpi), using anti-OsSAMS1 antibody. The red asterisk indicates the location of Pns11. (B) SAM …
Figure 6—figure supplement 1
GO biological process over-representation in differentially expressed genes after RDV infection and in transgenic rice plants.
(A) Venn diagrams showing the overlaps among differentially expressed genes after RDV infection and in OsSAMS1 (OsSAMS1 OX#25, Ossams1 KO#39) and S11 (S11 OX#11) transgenic rice plants. For …
Figure 7
Possible model for Pns11 enhancement of OsSAMS1 enzymatic activity to increase ethylene levels and enhance host susceptibility to viral infection.
The model proposes that the RDV-encoded protein Pns11 specifically interacts with OsSAMS1 to enhance its enzymatic activity, resulting in a corresponding increase in ethylene, and thus enhancing …
Additional files
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Supplementary file 1
Non-preference test.
- https://doi.org/10.7554/eLife.27529.025
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Supplementary file 2
Record of the number of rice plants showing RDV symptoms at time course and infection rates statistical analysis.
- https://doi.org/10.7554/eLife.27529.026
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Supplementary file 3
Differentially expressed genes of RDV-infected and OsSAMS1 and S11 transgenic rice RNA-seq analysis.
- https://doi.org/10.7554/eLife.27529.027
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Supplementary file 4
Responses of genes related to polyamine by RDV infection.
- https://doi.org/10.7554/eLife.27529.028
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Supplementary file 5
(A) Constructs in this study. (B) Primers for plasmids constructions in this study. (C) LC-MS/MS conditions. (D) Primers for RNA gel blot probes, real-time PCR, and semi-quantitative RT-PCR.
- https://doi.org/10.7554/eLife.27529.029
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Transparent reporting form
- https://doi.org/10.7554/eLife.27529.030
