{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T14:52:53Z","timestamp":1791298373127,"version":"4.1.0"},"reference-count":51,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"am","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000324","name":"Gatsby","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000324","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>In order to prevent damage by excess light, light-harvesting antennae can switch to an energy dissipative mode (termed non-photochemical quenching, NPQ). In higher plants, this switch is facilitated by the presence of photosystem II subunit S (PsbS) protein, which was discovered 25\u2005years ago. While the role of PsbS in induction of NPQ was soon found to require protonation of key glutamate residues facing the thylakoid lumen, a complete understanding of how NPQ is subsequently initiated is still lacking. Recent work on Norway spruce suggests that reversible phosphorylation at a few conserved residues of PsbS may affect its role in regulation of NPQ. Here we assessed PsbS phosphorylation changes in Arabidopsis and found these remained undetectable under control and combined chilling and high light stress conditions. We therefore used a genetic approach to assess potential functional implications of phosphorylation at threonine-259 (T259). Functional evaluation of point mutations at T259 in the background of PsbS knock-out mutant npq4 showed that neither phosphomimetic, nor phosphosubstitution, nor phosphonull substitutions could rescue NPQ activity to the level of the unperturbed protein, inconsistent with regulation via reversible phosphorylation. Instead, all residue substitutions at T259 gave rise to significantly impaired induction and accelerated NPQ recovery, while protein accumulation and thylakoid membrane localization were not affected. We suggest that these results point to a role for the C-terminus in the propensity or stability of hydrophobic interactions between PsbS and light-harvesting complex II antenna proteins to initiate the quenched state.<\/jats:p>","DOI":"10.1093\/jxb\/erag446","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T11:20:35Z","timestamp":1788780035000},"source":"Crossref","is-referenced-by-count":0,"title":["Mutation of a stromal C-terminal threonine residue of photosystem II subunit S slows down non-photochemical quenching induction and speeds up relaxation"],"prefix":"10.1093","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-9216-7139","authenticated-orcid":false,"given":"Weng Yik","family":"Chin","sequence":"first","affiliation":[{"name":"Department of Plant Sciences, University of Cambridge , Cambridge CB2 3EA ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6230-9626","authenticated-orcid":false,"given":"Julia","family":"Walter","sequence":"additional","affiliation":[{"name":"Department of Plant Sciences, University of Cambridge , Cambridge CB2 3EA ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4271-5587","authenticated-orcid":false,"given":"Alice K J","family":"Robijns","sequence":"additional","affiliation":[{"name":"Department of Plant Sciences, University of Cambridge , Cambridge CB2 3EA ,","place":["UK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4423-4100","authenticated-orcid":false,"given":"Johannes","family":"Kromdijk","sequence":"additional","affiliation":[{"name":"Department of Plant Sciences, University of Cambridge , Cambridge CB2 3EA ,","place":["UK"]},{"name":"Carl R Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign , Urbana, IL 61801 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"key":"2026100610162625900_erag446-B1","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate structure prediction of biomolecular interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"key":"2026100610162625900_erag446-B2","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1038\/s41477-020-0591-9","article-title":"An atypical short-chain dehydrogenase\u2013reductase functions in the relaxation of photoprotective qH in Arabidopsis","volume":"6","author":"Amstutz","year":"2020","journal-title":"Nature Plants"},{"key":"2026100610162625900_erag446-B3","doi-asserted-by":"crossref","first-page":"15265","DOI":"10.1073\/pnas.2533072100","article-title":"Light- and pH-dependent structural changes in the PsbS subunit of photosystem II","volume":"100","author":"Bergantino","year":"2003","journal-title":"Proceedings of the National Academy of Sciences, USA"},{"key":"2026100610162625900_erag446-B4","doi-asserted-by":"crossref","DOI":"10.1093\/gigascience\/giz126","article-title":"Access to RNA-sequencing data from 1,173 plant species: the 1000 Plant transcriptomes initiative (1KP)","volume":"8","author":"Carpenter","year":"2019","journal-title":"GigaScience"},{"key":"2026100610162625900_erag446-B5","doi-asserted-by":"crossref","DOI":"10.1016\/j.xplc.2024.101179","article-title":"Distinct features of PsbS essential for mediating plant photoprotection","volume":"6","author":"Chen","year":"2025","journal-title":"Plant Communications"},{"key":"2026100610162625900_erag446-B6","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1021\/acs.jpclett.2c03760","article-title":"pH-dependent conformational switch impacts stability of the PsbS dimer","volume":"14","author":"Chiariello","year":"2023","journal-title":"The Journal of Physical Chemistry Letters"},{"key":"2026100610162625900_erag446-B7","doi-asserted-by":"crossref","DOI":"10.1038\/nplants.2015.225","article-title":"Psbs interactions involved in the activation of energy dissipation in Arabidopsis","volume":"2","author":"Correa-Galvis","year":"2016","journal-title":"Nature Plants"},{"key":"2026100610162625900_erag446-B8","doi-asserted-by":"crossref","first-page":"17478","DOI":"10.1074\/jbc.M116.737312","article-title":"Photosystem II Subunit PsbS is involved in the induction of LHCSR protein-dependent energy dissipation in Chlamydomonas reinhardtii","volume":"291","author":"Correa-Galvis","year":"2016","journal-title":"The Journal of Biological Chemistry"},{"key":"2026100610162625900_erag446-B9","doi-asserted-by":"crossref","first-page":"2242","DOI":"10.1021\/bi500117c","article-title":"OGlcNAcylation and phosphorylation have similar structural effects in \u03b1-helices: post-translational modifications as inducible start and stop signals in \u03b1-helices, with greater structural effects on threonine modification","volume":"53","author":"Elbaum","year":"2014","journal-title":"Biochemistry"},{"key":"2026100610162625900_erag446-B10","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0035669","article-title":"Identification of key residues for pH dependent activation of violaxanthin de-epoxidase from Arabidopsis thaliana","volume":"7","author":"Fufezan","year":"2012","journal-title":"PLoS One"},{"key":"2026100610162625900_erag446-B11","doi-asserted-by":"crossref","first-page":"1747","DOI":"10.1104\/pp.15.00361","article-title":"In vivo identification of photosystem II light harvesting complexes interacting with PHOTOSYSTEM II SUBUNIT S","volume":"168","author":"Gerotto","year":"2015","journal-title":"Plant Physiology"},{"key":"2026100610162625900_erag446-B12","doi-asserted-by":"crossref","first-page":"D1178","DOI":"10.1093\/nar\/gkr944","article-title":"Phytozome: a comparative platform for green plant genomics","volume":"40","author":"Goodstein","year":"2012","journal-title":"Nucleic Acids Research"},{"key":"2026100610162625900_erag446-B13","doi-asserted-by":"crossref","first-page":"17499","DOI":"10.1073\/pnas.2004165117","article-title":"Specific thylakoid protein phosphorylations are prerequisites for overwintering of Norway spruce (Picea abies) photosynthesis","volume":"117","author":"Grebe","year":"2020","journal-title":"Proceedings of the National Academy of Sciences, USA"},{"key":"2026100610162625900_erag446-B14","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1042\/BJ20102155","article-title":"Optimized native gel systems for separation of thylakoid protein complexes: novel super- and mega-complexes","volume":"439","author":"J\u00e4rvi","year":"2011","journal-title":"The Biochemical Journal"},{"key":"2026100610162625900_erag446-B15","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1006\/abio.1994.1243","article-title":"A method for protein assay in Laemmli buffer","volume":"219","author":"Karlsson","year":"1994","journal-title":"Analytical Biochemistry"},{"key":"2026100610162625900_erag446-B16","first-page":"27","article-title":"Complementary PSII quantum yields calculated from simple fluorescence parameters measured by PAM fluorometry and the Saturation Pulse method","volume":"1","author":"Klughammer","year":"2008","journal-title":"PAM Application Notes"},{"key":"2026100610162625900_erag446-B17","doi-asserted-by":"crossref","DOI":"10.1038\/s41467-021-22530-4","article-title":"The molecular pH-response mechanism of the plant light-stress sensor PsbS","volume":"12","author":"Krishnan-Schmieden","year":"2021","journal-title":"Nature Communications"},{"key":"2026100610162625900_erag446-B18","doi-asserted-by":"crossref","DOI":"10.1111\/ppl.70298","article-title":"Moderate temperature reduction changes the high-light acclimation strategy of lettuce plants","volume":"177","author":"Lempi\u00e4inen","year":"2025","journal-title":"Physiologia Plantarum"},{"key":"2026100610162625900_erag446-B19","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1038\/35000131","article-title":"A pigment-binding protein essential for regulation of photosynthetic light harvesting","volume":"403","author":"Li","year":"2000","journal-title":"Nature"},{"key":"2026100610162625900_erag446-B20","doi-asserted-by":"crossref","first-page":"33590","DOI":"10.1074\/jbc.M204797200","article-title":"Molecular and global time-resolved analysis of a psbS gene dosage effect on pH- and xanthophyll cycle-dependent nonphotochemical quenching in photosystem II","volume":"277","author":"Li","year":"2002","journal-title":"The Journal of Biological Chemistry"},{"key":"2026100610162625900_erag446-B21","doi-asserted-by":"crossref","first-page":"15222","DOI":"10.1073\/pnas.232447699","article-title":"PsbS-dependent enhancement of feedback de-excitation protects photosystem II from photoinhibition","volume":"99","author":"Li","year":"2002","journal-title":"Proceedings of the National Academy of Sciences, USA"},{"key":"2026100610162625900_erag446-B22","doi-asserted-by":"crossref","first-page":"22866","DOI":"10.1074\/jbc.M402461200","article-title":"Regulation of photosynthetic light harvesting involves intrathylakoid lumen pH sensing by the PsbS protein","volume":"279","author":"Li","year":"2004","journal-title":"The Journal of Biological Chemistry"},{"key":"2026100610162625900_erag446-B23","doi-asserted-by":"crossref","first-page":"1737","DOI":"10.1021\/acs.jpclett.9b00437","article-title":"Molecular anatomy of plant photoprotective switches: the sensitivity of PsbS to the environment, residue by residue","volume":"10","author":"Liguori","year":"2019","journal-title":"The Journal of Physical Chemistry Letters"},{"key":"2026100610162625900_erag446-B24","doi-asserted-by":"crossref","DOI":"10.21105\/joss.01541","article-title":"Bayestestr: describing effects and their uncertainty, existence and significance within the Bayesian framework","volume":"4","author":"Makowski","year":"2019","journal-title":"Journal of Open Source Software"},{"key":"2026100610162625900_erag446-B25","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.envexpbot.2018.05.005","article-title":"Photoinhibition or photoprotection of photosynthesis? Update on the (newly termed) sustained quenching component qH","volume":"154","author":"Malno\u00eb","year":"2018","journal-title":"Environmental and Experimental Botany"},{"key":"2026100610162625900_erag446-B26","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1093\/treephys\/tpx065","article-title":"A comparison of pine and spruce in recovery from winter stress; changes in recovery kinetics, and the abundance and phosphorylation status of photosynthetic proteins during winter","volume":"37","author":"Merry","year":"2017","journal-title":"Tree Physiology"},{"key":"2026100610162625900_erag446-B27","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1016\/j.molcel.2011.02.011","article-title":"Threonine 22 phosphorylation attenuates Hsp90 interaction with cochaperones and affects its chaperone activity","volume":"41","author":"Mollapour","year":"2012","journal-title":"Molecular Cell"},{"key":"2026100610162625900_erag446-B28","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1002\/psc.310010507","article-title":"Relationship of sidechain hydrophobicity and \u03b1-helical propensity on the stability of the single-stranded amphipathic \u03b1-helix","volume":"1","author":"Monera","year":"1995","journal-title":"Journal of Peptide Science"},{"key":"2026100610162625900_erag446-B29","doi-asserted-by":"crossref","first-page":"970","DOI":"10.1104\/pp.010924","article-title":"Ascorbate deficiency can limit violaxanthin de-epoxidase activity in vivo","volume":"128","author":"M\u00fcller-Moul\u00e9 P Conklin","year":"2002","journal-title":"Plant Physiology"},{"key":"2026100610162625900_erag446-B30","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1016\/j.bbabio.2010.01.001","article-title":"Identification of a slowly inducible zeaxanthin-dependent component of non-photochemical quenching of chlorophyll fluorescence generated under steady-state conditions in Arabidopsis","volume":"1797","author":"Nilkens","year":"2010","journal-title":"Biochimica et Biophysica Acta \u2013 Bioenergetics"},{"key":"2026100610162625900_erag446-B31","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.1105\/tpc.10.7.1121","article-title":"Arabidopsis mutants define a central role for the xanthophyll cycle in the regulation of photosynthetic energy conversion","volume":"10","author":"Niyogi","year":"1998","journal-title":"The Plant Cell"},{"key":"2026100610162625900_erag446-B32","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1093\/jxb\/eri056","article-title":"Is PsbS the site of non-photochemical quenching in photosynthesis?","volume":"56","author":"Niyogi","year":"2005","journal-title":"Journal of Experimental Botany"},{"key":"2026100610162625900_erag446-B33","doi-asserted-by":"crossref","DOI":"10.3389\/fpls.2015.00562","article-title":"Functional analysis of the BRI1 receptor kinase by Thr-for-Ser substitution in a regulatory autophosphorylation site","volume":"6","author":"Oh","year":"2015","journal-title":"Frontiers in Plant Science"},{"key":"2026100610162625900_erag446-B34","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1038\/s41586-019-1693-2","article-title":"One thousand plant transcriptomes and the phylogenomics of green plants","volume":"574","author":"One Thousand Plant Transcriptomes Initiative","year":"2019","journal-title":"Nature"},{"key":"2026100610162625900_erag446-B35","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1146\/annurev.arplant.54.072402.115741","article-title":"Photosynthesis of overwintering evergreen plants","volume":"54","author":"\u00d6quist","year":"2003","journal-title":"Annual Review of Plant Biology"},{"key":"2026100610162625900_erag446-B36","author":"Pag\u00e8s","year":"2024"},{"key":"2026100610162625900_erag446-B37","doi-asserted-by":"crossref","first-page":"1938","DOI":"10.1021\/acschembio.3c00068","article-title":"An inherent difference between serine and threonine phosphorylation: phosphothreonine strongly prefers a highly ordered, compact, cyclic conformation","volume":"18","author":"Pandey","year":"2023","journal-title":"ACS Chemical Biology"},{"key":"2026100610162625900_erag446-B38","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1007\/s11120-020-00740-z","article-title":"On the PsbS-induced quenching in the plant major light-harvesting complex LHCII studied in proteoliposomes","volume":"144","author":"Pawlak","year":"2020","journal-title":"Photosynthesis Research"},{"key":"2026100610162625900_erag446-B39","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1039\/d0pp00025f","article-title":"Composition, phosphorylation and dynamic organization of photosynthetic protein complexes in plant thylakoid membrane","volume":"19","author":"Rantala","year":"2020","journal-title":"Photochemical & Photobiological Sciences"},{"key":"2026100610162625900_erag446-B40","doi-asserted-by":"crossref","DOI":"10.1016\/j.bbabio.2020.148183","article-title":"Psbs contributes to photoprotection in Chlamydomonas reinhardtii independently of energy dissipation","volume":"1861","author":"Redekop","year":"2020","journal-title":"Biochimica et Biophysica Acta \u2013 Bioenergetics"},{"key":"2026100610162625900_erag446-B41","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1074\/mcp.M114.040352","article-title":"Quantitative phosphoproteomics of the Ataxia Telangiectasia-Mutated (ATM) and Ataxia Telangiectasia-Mutated and Rad3-related (ATR) dependent DNA damage response in Arabidopsis thaliana","volume":"14","author":"Roitinger","year":"2015","journal-title":"Molecular & Cellular Proteomics"},{"key":"2026100610162625900_erag446-B42","doi-asserted-by":"crossref","first-page":"1903","DOI":"10.1104\/pp.15.01935","article-title":"Nonphotochemical chlorophyll fluorescence quenching: mechanism and effectiveness in protecting plants from photodamage","volume":"170","author":"Ruban","year":"2016","journal-title":"Plant Physiology"},{"key":"2026100610162625900_erag446-B43","doi-asserted-by":"crossref","DOI":"10.1038\/nplants.2016.225","article-title":"The xanthophyll cycle affects reversible interactions between PsbS and light-harvesting complex II to control non-photochemical quenching","volume":"3","author":"Sacharz","year":"2017","journal-title":"Nature Plants"},{"key":"2026100610162625900_erag446-B44","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1111\/j.1365-313X.2009.04060.x","article-title":"A rapid and non-destructive screenable marker, FAST, for identifying transformed seeds of Arabidopsis thaliana","volume":"61","author":"Shimada","year":"2010","journal-title":"The Plant Journal"},{"key":"2026100610162625900_erag446-B45","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/978-1-0716-2079-3_11","volume-title":"Plant proteases and plant cell death. Methods in Molecular Biology","author":"Stael","year":"2022"},{"key":"2026100610162625900_erag446-B46","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1038\/nrm2203","article-title":"Mechanisms of specificity in protein phosphorylation","volume":"8","author":"Ubersax","year":"2007","journal-title":"Nature Reviews: Molecular Cell Biology"},{"key":"2026100610162625900_erag446-B47","doi-asserted-by":"crossref","first-page":"33679","DOI":"10.1074\/jbc.M505729200","article-title":"STN8 protein kinase in Arabidopsis thaliana is specific in phosphorylation of photosystem II core proteins","volume":"280","author":"Vainonen","year":"2005","journal-title":"The Journal of Biological Chemistry"},{"key":"2026100610162625900_erag446-B48","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0016765","article-title":"A modular cloning system for standardized assembly of multigene constructs","volume":"6","author":"Weber","year":"2011","journal-title":"PLoS One"},{"key":"2026100610162625900_erag446-B49","doi-asserted-by":"crossref","DOI":"10.1201\/9781003279242","volume-title":"Data integration, manipulation and visualization of phylogenetic trees","author":"Yu","year":"2022","edition":":"},{"key":"2026100610162625900_erag446-B50","doi-asserted-by":"crossref","first-page":"15757","DOI":"10.1073\/pnas.1211017109","article-title":"A kinetic model of rapidly reversible nonphotochemical quenching","volume":"109","author":"Zaks","year":"2012","journal-title":"Proceedings of the National Academy of Sciences, USA"},{"key":"2026100610162625900_erag446-B51","doi-asserted-by":"crossref","DOI":"10.1093\/bib\/bbac222","article-title":"Ggmsa: a visual exploration tool for multiple sequence alignment and associated data","volume":"23","author":"Zhou","year":"2022","journal-title":"Briefings in Bioinformatics"}],"container-title":["Journal of Experimental Botany"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jxb\/advance-article-pdf\/doi\/10.1093\/jxb\/erag446\/70989504\/erag446.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jxb\/advance-article-pdf\/doi\/10.1093\/jxb\/erag446\/70989504\/erag446.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T14:30:41Z","timestamp":1791297041000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jxb\/advance-article\/doi\/10.1093\/jxb\/erag446\/8787795"}},"subtitle":[],"editor":[{"given":"Tracy","family":"Lawson","sequence":"additional","affiliation":[{"name":"University of Illinois Urbana-Champaign ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,8]]},"references-count":51,"URL":"https:\/\/doi.org\/10.1093\/jxb\/erag446","relation":{},"ISSN":["0022-0957","1460-2431"],"issn-type":[{"value":"0022-0957","type":"print"},{"value":"1460-2431","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,8]]},"article-number":"erag446"}}