The xanthophyll cycle affects reversible interactions between PsbS and light-harvesting complex II to control non-photochemical quenching

Abstract

To maintain high photosynthetic rates, plants must adapt to their light environment on a timescale of seconds to minutes. Therefore, the light-harvesting antenna system of photosystem II in thylakoid membranes, light-harvesting complex II (LHCII), has a feedback mechanism, which determines the proportion of absorbed energy dissipated as heat: non-photochemical chlorophyll fluorescence quenching (NPQ). This is crucial to prevent photo-oxidative damage to photosystem II (PSII) and is controlled by the transmembrane pH differences (ΔpH). High ΔpH activates NPQ by protonation of the protein PsbS and the enzymatic de-epoxidation of LHCII-bound violaxanthin to zeaxanthin. But the precise role of PsbS and its interactions with different LHCII complexes remain uncertain. We have investigated PsbS–LHCII interactions in native thylakoid membranes using magnetic-bead-linked antibody pull-downs. The interaction of PsbS with the antenna system is affected by both ΔpH and the level of zeaxanthin. In the presence of ΔpH alone, PsbS is found to be mainly associated with the trimeric LHCII protein polypeptides, Lhcb1, Lhcb2 and Lhcb3. However, a combination of ΔpH and zeaxanthin increases the proportion of PsbS bound to the minor LHCII antenna complex proteins Lhcb4, Lhcb5 and Lhcb6. This pattern of interaction is not influenced by the presence of PSII reactions centres. Similar to LHCII particles in the photosynthetic membrane, PsbS protein forms clusters in the NPQ state. NPQ recovery in the dark requires uncoupling of PsbS. We suggest that PsbS acts as a ‘seeding’ centre for the LHCII antenna rearrangement that is involved in NPQ.

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Figure 1: Affinity pull-down assay on violaxanthin-containing spinach membranes.
Figure 2: Affinity pull-down assay on zeaxanthin-containing spinach membranes.
Figure 3: Fast protein liquid chromatography (FPLC) gel-filtration separation of A. thaliana L17 transgenic line membranes in the dark and NPQ state.
Figure 4: Localization of PsbS in dark-adapted and light-treated spinach chloroplasts based on electron microscopy of immunogold-labelled negatively stained thin sections of the fixed material.
Figure 5: Model of PsbS interactions within the PSII–LHCII complex.

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Acknowledgements

A.V.R. acknowledges funding from the Biotechnology and Biological Sciences Research Council of the UK and the Leverhulme Trust. The authors acknowledge Y. Tian for the help with plant growth, as well as M. Johnson and C. Duffy for critical reading of the manuscript.

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J.S. and A.V.R. designed the experiments. J.S. performed biochemistry, PAM fluorescence measurements, preparation for TEM and particle analysis. V.G. performed PAM fluorescence measurements. P.U. assisted with the experiments. G.M. operated TEM and assisted with sample preparation. All authors discussed the results and commented on the manuscript. J.S., V.G. and A.V.R. wrote the manuscript.

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Correspondence to Alexander V. Ruban.

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The authors declare no competing financial interests.

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Sacharz, J., Giovagnetti, V., Ungerer, P. et al. The xanthophyll cycle affects reversible interactions between PsbS and light-harvesting complex II to control non-photochemical quenching. Nature Plants 3, 16225 (2017). https://doi.org/10.1038/nplants.2016.225

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