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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.


Subpopulations of FLS2 and BRI1 co-localise with BAK1 in plasma membrane nanodomains.The images show the plasma membrane localisation of FLS2-GFP or BRI1-GFP with BAK1- mCherry. The white arrowheads indicate plasma membrane nanodomains that contain fluorescently labelled receptor and co-receptor molecules simultaneously. The images were acquired 5 days post germination using VAEM and the scale bars represent a distance of 5 µm. The lines were generated by crossing pFLS2::FLS2-GFP (Göhre et al., 2008) or pBRI1::BRI1- GFP (Friedrichsen et al., 2000), respectively, with pBAK1::BAK1-mCherry (Bücherl et al., 2013).DOI:http://dx.doi.org/10.7554/eLife.25114.023
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fig10: Subpopulations of FLS2 and BRI1 co-localise with BAK1 in plasma membrane nanodomains.The images show the plasma membrane localisation of FLS2-GFP or BRI1-GFP with BAK1- mCherry. The white arrowheads indicate plasma membrane nanodomains that contain fluorescently labelled receptor and co-receptor molecules simultaneously. The images were acquired 5 days post germination using VAEM and the scale bars represent a distance of 5 µm. The lines were generated by crossing pFLS2::FLS2-GFP (Göhre et al., 2008) or pBRI1::BRI1- GFP (Friedrichsen et al., 2000), respectively, with pBAK1::BAK1-mCherry (Bücherl et al., 2013).DOI:http://dx.doi.org/10.7554/eLife.25114.023

Mentions: We nevertheless provide here some preliminary evidence for the co-localisation of BAK1- mCherry with FLS2-GFP and BRI1-GFP, respectively, within PM nanodomains of stable transgenic Arabidopsis lines (see Author response image 1). Moreover, data included in a manuscript submitted elsewhere show BRI1/BAK1 association within such nanodomains (Hutten et al., under revision with PLoS One; co-last authors: Johannes Hohlbein & Jan Willem Borst; co-author: CA Bücherl). Unfortunately, we have been so far not able to address this question for the FLS2/BAK1 combination, due to technical feasibility regarding FLIM in living leaf tissue and our inability to detect red fluorescent when expressing FLS2-RFP in stable transgenic plants.10.7554/eLife.25114.023Author response image 1.Subpopulations of FLS2 and BRI1 co-localise with BAK1 in plasma membrane nanodomains.


Plant immune and growth receptors share common signalling components but localise to distinct plasma membrane nanodomains
Subpopulations of FLS2 and BRI1 co-localise with BAK1 in plasma membrane nanodomains.The images show the plasma membrane localisation of FLS2-GFP or BRI1-GFP with BAK1- mCherry. The white arrowheads indicate plasma membrane nanodomains that contain fluorescently labelled receptor and co-receptor molecules simultaneously. The images were acquired 5 days post germination using VAEM and the scale bars represent a distance of 5 µm. The lines were generated by crossing pFLS2::FLS2-GFP (Göhre et al., 2008) or pBRI1::BRI1- GFP (Friedrichsen et al., 2000), respectively, with pBAK1::BAK1-mCherry (Bücherl et al., 2013).DOI:http://dx.doi.org/10.7554/eLife.25114.023
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Related In: Results  -  Collection

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fig10: Subpopulations of FLS2 and BRI1 co-localise with BAK1 in plasma membrane nanodomains.The images show the plasma membrane localisation of FLS2-GFP or BRI1-GFP with BAK1- mCherry. The white arrowheads indicate plasma membrane nanodomains that contain fluorescently labelled receptor and co-receptor molecules simultaneously. The images were acquired 5 days post germination using VAEM and the scale bars represent a distance of 5 µm. The lines were generated by crossing pFLS2::FLS2-GFP (Göhre et al., 2008) or pBRI1::BRI1- GFP (Friedrichsen et al., 2000), respectively, with pBAK1::BAK1-mCherry (Bücherl et al., 2013).DOI:http://dx.doi.org/10.7554/eLife.25114.023
Mentions: We nevertheless provide here some preliminary evidence for the co-localisation of BAK1- mCherry with FLS2-GFP and BRI1-GFP, respectively, within PM nanodomains of stable transgenic Arabidopsis lines (see Author response image 1). Moreover, data included in a manuscript submitted elsewhere show BRI1/BAK1 association within such nanodomains (Hutten et al., under revision with PLoS One; co-last authors: Johannes Hohlbein & Jan Willem Borst; co-author: CA Bücherl). Unfortunately, we have been so far not able to address this question for the FLS2/BAK1 combination, due to technical feasibility regarding FLIM in living leaf tissue and our inability to detect red fluorescent when expressing FLS2-RFP in stable transgenic plants.10.7554/eLife.25114.023Author response image 1.Subpopulations of FLS2 and BRI1 co-localise with BAK1 in 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.