Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

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.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

References

  1. Demmig-Adams, B., Garab, G., Adams, W. III & Govindjee . Non-Photochemical Quenching and Energy Dissipation in Plants, Algae and Cyanobacteria (Springer, 2014).

    Book  Google Scholar 

  2. Jansson, S. A guide to the Lhc genes and their relatives in Arabidopsis. Trends Plant Sci. 4, 236–240 (1999).

    Article  CAS  Google Scholar 

  3. Powles, S. B. Photoinhibition of photosynthesis induced by visible-light. Annu. Rev. Plant Physiol. Plant Mol. Biol. 35, 15–44 (1984).

    Article  CAS  Google Scholar 

  4. Barber, J. Molecular basis of the vulnerability of photosystem II to damage by light. Aust. J. Plant Physiol. 22, 201–208 (1995).

    CAS  Google Scholar 

  5. Osmond, C. B. in Photoinhibition of Photosynthesis (eds Baker, N. R. & Bowyer, J. R. ) 1–24 (Bios Scientific Publishers, 1994).

    Google Scholar 

  6. Demmig-Adams, B. & Adams, W. W. Photoprotection and other responses of plants to high light stress. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43, 599–626 (1992).

    Article  CAS  Google Scholar 

  7. Li, Z., Wakao, S., Fischer, B. B. & Niyogi, K. K. Sensing and responding to excess light. Annu. Rev. Plant Biol. 60, 239–260 (2009).

    Article  CAS  Google Scholar 

  8. Horton, P., Ruban, A. V. & Walters, R. G. Regulation of light harvesting in green plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47, 655–684 (1996).

    Article  CAS  Google Scholar 

  9. Niyogi, K. K. & Truong, T. B. Evolution of flexible non-photochemical quenching mechanisms that regulate light harvesting in oxygenic photosynthesis. Curr. Opin. Plant Biol. 16, 307–314 (2013).

    Article  CAS  Google Scholar 

  10. Ruban, A. V. Evolution under the sun: optimizing light harvesting in photosynthesis. J. Exp. Bot. 66, 7–23 (2015).

    Article  CAS  Google Scholar 

  11. Ruban, A. V. Non-photochemical chlorophyll fluorescence quenching: mechanism and effectiveness in protection against photodamage. Plant Physiol. 170, 1903–1916 (2016).

    Article  CAS  Google Scholar 

  12. Kramer, D. M., Sacksteder, C. A. & Cruz, J. A. How acidic is the lumen? Photosynth. Res. 60, 151–163 (1999).

    Article  CAS  Google Scholar 

  13. Liu, Z. et al. Crystal structure of spinach major light-harvesting complex at 2.72 Å resolution. Nature 428, 287–229 (2004).

    Article  CAS  Google Scholar 

  14. Demmig-Adams, B. Carotenoids and photoprotection: a role for the xanthophyll zeaxanthin. Biochim. Biophys. Acta 1020, 1–2 (1990).

    Article  CAS  Google Scholar 

  15. Yamamoto, H. Y., Nakayama, T. O. M. & Chichester, C. O. Studies on the light and dark interconversions of leaf xanthophylls. Arch. Biochem. Biophys. 97, 168–173 (1962).

    Article  CAS  Google Scholar 

  16. Li, X. P. et al. A pigment-binding protein essential for regulation of photosynthetic light harvesting. Nature 403, 391–395 (2000).

    Article  CAS  Google Scholar 

  17. Li, X. P. et al. Regulation of photosynthetic light harvesting involves intrathylakoid lumen pH sensing by the PsbS protein. J. Biol. Chem. 279, 22866–22287 (2004).

    Article  CAS  Google Scholar 

  18. Fan, M. et al. Crystal structures of the PsbS protein essential for photoprotection. Nat. Struc. Mol. Biol. 22, 729–735 (2015).

    Article  CAS  Google Scholar 

  19. Bergantino, E. et al. Light- and pH-dependent structural changes in the PsbS protein of photosystem II. Proc. Natl Acad. Sci. USA 100, 15265–15270 (2003).

    Article  CAS  Google Scholar 

  20. Teardo, E. et al. Evidences for interaction of PsbS with photosynthetic complexes in maize thylakoids. Biochim. Biophys. Acta 1767, 703–711 (2007).

    Article  CAS  Google Scholar 

  21. Wilk, L., Grunwald, M., Liao, P. N., Walla, P. J. & Kühlbrandt, W. Direct interaction of the major light-harvesting complex II and PsbS in nonphotochemical quenching. Proc. Natl Acad. Sci. USA 110, 5452–5456 (2013).

    Article  CAS  Google Scholar 

  22. Gerotto, C., Franchin, C., Arrigoni, G. & Morosinotto, T. In vivo identification of photosystem II light harvesting complexes interacting with photosystem II subunit S. Plant Physiol. 168, 1747–1761 (2015).

    Article  CAS  Google Scholar 

  23. Correa-Galvis, V., Poschmann, G., Melzer, M., Stühler, K. & Jahns, P. PsbS interactions involved in the activation of energy dissipation in Arabidopsis. Nat. Plants. 2, 15225 (2016).

    Article  CAS  Google Scholar 

  24. Albanese, P. et al. Dynamic reorganization of photosystem II supercomplexes in response to variations in light intensities. Biochim. Biophys. Acta. 10, 651–660 (2016).

    Google Scholar 

  25. Johnson, M. P. & Ruban, A. V. Arabidopsis plants lacking PsbS protein possess photoprotective energy dissipation. Plant J. 61, 283–289 (2010).

    Article  CAS  Google Scholar 

  26. Zia, A., Johnson, M. P. & Ruban, A. V. Acclimation- and mutation-induced enhancement of PsbS levels affects the kinetics of nonphotochemical quenching in Arabidopsis thaliana. Planta 233, 1253–1264 (2011).

    Article  CAS  Google Scholar 

  27. Belgio, E., Johnson, M. P., Jurić, S. & Ruban, A. V. Higher plant photosystem II light-harvesting antenna, not the reaction center, determines the excited-state lifetime – both the maximum and the nonphotochemically quenched. Biophys. J. 102, 2761–2771 (2012).

    Article  CAS  Google Scholar 

  28. Belgio, E., Ungerer, P. & Ruban, A. V. Light-harvesting superstructures of green plant chloroplasts lacking photosystems. Plant Cell Environ. 38, 2035–2047 (2015).

    Article  CAS  Google Scholar 

  29. Ilioaia, C., Duffy, C. D. P. & Ruban, A. V. Changes in the energy transfer pathways within photosystem II antenna induced by the xanthophyll cycle activity. J. Phys. Chem. B 117, 5841–5847 (2013).

    Article  CAS  Google Scholar 

  30. Johnson, M. P. et al. Photoprotective energy dissipation involves the reorganization of photosystem II light-harvesting complexes in the grana membranes of spinach chloroplasts. Plant Cell 23, 1468–1479 (2011).

    Article  CAS  Google Scholar 

  31. Ware, M. A., Giovagnetti, V., Belgio, E. & Ruban, A. V. PsbS protein modulates non-photochemical chlorophyll fluorescence quenching in membranes depleted from photosystems. J. Photochem. Photobiol. B 152, 301–307 (2015).

    Article  CAS  Google Scholar 

  32. Crouchman, S., Ruban, A. & Horton, P. PsbS enhances nonphotochemical fluorescence quenching in the absence of zeaxanthin. FEBS Lett. 580, 2053–2058 (2006).

    Article  CAS  Google Scholar 

  33. Kereïche, S., Kiss, A. Z., Kouril, R., Boekema, E. & Horton, P. The PsbS protein controls the macro-organisation of photosystem II complexes in the grana membranes of higher plant chloroplasts. FEBS Lett. 584, 754–764 (2010).

    Article  Google Scholar 

  34. Goral, T. et al. Light-harvesting antenna composition controls the macrostructure and dynamics of thylakoid membranes in Arabidopsis. Plant J. 69, 289–301 (2012).

    Article  CAS  Google Scholar 

  35. Porra, R. J., Thompson, W. A. & Kriedemann, E. E. Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim. Biophys. Acta. 975, 384–394 (1989).

    Article  CAS  Google Scholar 

  36. Ruban, A. V. et al. Plasticity in the composition of the light harvesting antenna of higher plants preserves structural integrity and biological function. J. Biol. Chem. 281, 14981–14990 (2006).

    Article  CAS  Google Scholar 

  37. Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970).

    Article  CAS  Google Scholar 

  38. Kouřil, R., Dekker, J. P. & Boekema, E. J. Supramolecular structure of photosystem II in green plants. Biochim. Biophys. Acta 1817, 2–12 (2012).

    Article  Google Scholar 

Download references

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.

Author information

Authors and Affiliations

Authors

Contributions

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.

Corresponding author

Correspondence to Alexander V. Ruban.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Figures 1–5. (PDF 923 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1038/nplants.2016.225

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing