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Plant immune and growth receptors share common signalling components but localise to distinct plasma membrane nanodomains

View Article: PubMed Central - PubMed

ABSTRACT

Cell surface receptors govern a multitude of signalling pathways in multicellular organisms. In plants, prominent examples are the receptor kinases FLS2 and BRI1, which activate immunity and steroid-mediated growth, respectively. Intriguingly, despite inducing distinct signalling outputs, both receptors employ common downstream signalling components, which exist in plasma membrane (PM)-localised protein complexes. An important question is thus how these receptor complexes maintain signalling specificity. Live-cell imaging revealed that FLS2 and BRI1 form PM nanoclusters. Using single-particle tracking we could discriminate both cluster populations and we observed spatiotemporal separation between immune and growth signalling platforms. This finding was confirmed by visualising FLS2 and BRI1 within distinct PM nanodomains marked by specific remorin proteins and differential co-localisation with the cytoskeleton. Our results thus suggest that signalling specificity between these pathways may be explained by the spatial separation of FLS2 and BRI1 with their associated signalling components within dedicated PM nanodomains.

Doi:: http://dx.doi.org/10.7554/eLife.25114.001

No MeSH data available.


Control experiments for verifying the specific co-localisation of FLS2 and BRI1 with remorin markers.The coloured data points indicate the values of technical replicates; black dots indicate the position of outliers. To assess whether the determined co-localisation values (orig.) were significant, co-localisation analysis was also carried out after image randomisation. For this purpose, one of the two image channels was rotated by 90 degrees prior to co-localisation analysis (rota.). As shown by the represented p-values, FLS2-GFP (FLS2) showed to a varying degree specific co-localisation with the four tested mRFP-tagged remorin markers. In contrast, BRI1-GFP (BRI1) only specifically co-localised with mRFP-REM6.1 (REM6.1) and mRFP-REM6.2 (REM6.2). The indicated p-values were obtained using a two-tailed heteroscedastic t-test and a Bonferroni multiple hypothesis correction.DOI:http://dx.doi.org/10.7554/eLife.25114.018
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fig7s1: Control experiments for verifying the specific co-localisation of FLS2 and BRI1 with remorin markers.The coloured data points indicate the values of technical replicates; black dots indicate the position of outliers. To assess whether the determined co-localisation values (orig.) were significant, co-localisation analysis was also carried out after image randomisation. For this purpose, one of the two image channels was rotated by 90 degrees prior to co-localisation analysis (rota.). As shown by the represented p-values, FLS2-GFP (FLS2) showed to a varying degree specific co-localisation with the four tested mRFP-tagged remorin markers. In contrast, BRI1-GFP (BRI1) only specifically co-localised with mRFP-REM6.1 (REM6.1) and mRFP-REM6.2 (REM6.2). The indicated p-values were obtained using a two-tailed heteroscedastic t-test and a Bonferroni multiple hypothesis correction.DOI:http://dx.doi.org/10.7554/eLife.25114.018

Mentions: Based on the findings of Keinath et al. (2010), we investigated the spatial relationship between the two LRR-RKs and REM1.2 in leaf epidermal cells. As shown in Figure 7A I–A III and I, we identified a positive correlation for FLS2-GFP and mRFP-REM1.2 fluorescence intensities. In contrast, the co-localisation of this PM nanodomain marker with BRI1 was unspecific (Figure 7B I−B III and I, Figure 7—figure supplement 1). Similar results were obtained using mRFP-REM1.3 as PM nanodomain marker (Figure 7C I–D III and I, Figure 7—figure supplement 1).10.7554/eLife.25114.017Figure 7.FLS2 and BRI1 co-localize differentially with remorin markers.


Plant immune and growth receptors share common signalling components but localise to distinct plasma membrane nanodomains
Control experiments for verifying the specific co-localisation of FLS2 and BRI1 with remorin markers.The coloured data points indicate the values of technical replicates; black dots indicate the position of outliers. To assess whether the determined co-localisation values (orig.) were significant, co-localisation analysis was also carried out after image randomisation. For this purpose, one of the two image channels was rotated by 90 degrees prior to co-localisation analysis (rota.). As shown by the represented p-values, FLS2-GFP (FLS2) showed to a varying degree specific co-localisation with the four tested mRFP-tagged remorin markers. In contrast, BRI1-GFP (BRI1) only specifically co-localised with mRFP-REM6.1 (REM6.1) and mRFP-REM6.2 (REM6.2). The indicated p-values were obtained using a two-tailed heteroscedastic t-test and a Bonferroni multiple hypothesis correction.DOI:http://dx.doi.org/10.7554/eLife.25114.018
© Copyright Policy
Related In: Results  -  Collection

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fig7s1: Control experiments for verifying the specific co-localisation of FLS2 and BRI1 with remorin markers.The coloured data points indicate the values of technical replicates; black dots indicate the position of outliers. To assess whether the determined co-localisation values (orig.) were significant, co-localisation analysis was also carried out after image randomisation. For this purpose, one of the two image channels was rotated by 90 degrees prior to co-localisation analysis (rota.). As shown by the represented p-values, FLS2-GFP (FLS2) showed to a varying degree specific co-localisation with the four tested mRFP-tagged remorin markers. In contrast, BRI1-GFP (BRI1) only specifically co-localised with mRFP-REM6.1 (REM6.1) and mRFP-REM6.2 (REM6.2). The indicated p-values were obtained using a two-tailed heteroscedastic t-test and a Bonferroni multiple hypothesis correction.DOI:http://dx.doi.org/10.7554/eLife.25114.018
Mentions: Based on the findings of Keinath et al. (2010), we investigated the spatial relationship between the two LRR-RKs and REM1.2 in leaf epidermal cells. As shown in Figure 7A I–A III and I, we identified a positive correlation for FLS2-GFP and mRFP-REM1.2 fluorescence intensities. In contrast, the co-localisation of this PM nanodomain marker with BRI1 was unspecific (Figure 7B I−B III and I, Figure 7—figure supplement 1). Similar results were obtained using mRFP-REM1.3 as PM nanodomain marker (Figure 7C I–D III and I, Figure 7—figure supplement 1).10.7554/eLife.25114.017Figure 7.FLS2 and BRI1 co-localize differentially with remorin markers.

View Article: PubMed Central - PubMed

ABSTRACT

Cell surface receptors govern a multitude of signalling pathways in multicellular organisms. In plants, prominent examples are the receptor kinases FLS2 and BRI1, which activate immunity and steroid-mediated growth, respectively. Intriguingly, despite inducing distinct signalling outputs, both receptors employ common downstream signalling components, which exist in plasma membrane (PM)-localised protein complexes. An important question is thus how these receptor complexes maintain signalling specificity. Live-cell imaging revealed that FLS2 and BRI1 form PM nanoclusters. Using single-particle tracking we could discriminate both cluster populations and we observed spatiotemporal separation between immune and growth signalling platforms. This finding was confirmed by visualising FLS2 and BRI1 within distinct PM nanodomains marked by specific remorin proteins and differential co-localisation with the cytoskeleton. Our results thus suggest that signalling specificity between these pathways may be explained by the spatial separation of FLS2 and BRI1 with their associated signalling components within dedicated PM nanodomains.

Doi:: http://dx.doi.org/10.7554/eLife.25114.001

No MeSH data available.