The photosynthetic machinery of plants can acclimate to changes in light conditions by balancing light-harvesting between the two photosystems (PS). This acclimation response is induced by the change in the redox state of the plastoquinone pool, which triggers state transitions through activation of the STN7 kinase and subsequent phosphorylation of light-harvesting complex II (LHCII) proteins. Phosphorylation of LHCII results in its association with PSI (state 2), whereas dephosphorylation restores energy allocation to PSII (state 1). In addition to state transition regulation by phosphorylation, we have recently discovered that plants lacking the chloroplast acetyltransferase NSI are also locked in state 1, even though they possess normal LHCII phosphorylation. This defect may result from decreased lysine acetylation of several chloroplast proteins. Here, we compared the composition of wild type (wt), stn7 and nsi thylakoid protein complexes involved in state transitions separated by Blue Native gel electrophoresis. Protein complex composition and relative protein abundances were determined by LC-MS/MS analyses using iBAQ quantification. We show that despite obvious mechanistic differences leading to defects in state transitions, no major differences were detected in the composition of PSI and LHCII between the mutants. Moreover, both stn7 and nsi plants show retarded growth and decreased PSII capacity under fluctuating light as compared to wt, while the induction of non-photochemical quenching under fluctuating light was much lower in both nsi mutants than in stn7.

Centre Algatech Institute of Microbiology Czech Academy of Sciences Novohradská 237 Opatovický mlýn 379 81 Třebon Czech Republic

Compact Plants Phenomics Center Washington State University Pullman WA 99164 USA

Department of Biochemistry Molecular Plant Biology University of Turku Biocity A Tykistökatu 6 20520 Turku Finland

Institute of Biological Chemistry Washington State University Pullman WA 99164 6340 USA

Plant Physiology Institute of Plant Biology and Biotechnology University of Münster Schlossplatz 7 48149 Münster Germany

Plant Physiology School of Biological Sciences Washington State University Pullman WA 99164 4236 USA

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Adamska I, Roobol-Boza M, Lindahl M, Andersson B. Isolation of pigment-binding early light-inducible proteins from pea. Eur J Biochem. 1999;260:453–460. PubMed

Albertsson P. A quantitative model of the domain structure of the photosynthetic membrane. Trends Plant Sci. 2001;6:349–358. PubMed

Allahverdiyeva Y, Mamedov F, Holmström M, Nurmi M, Lundin B, Styring S, Spetea C, Aro EM. Comparison of the electron transport properties of the psbo1 and psbo2 mutants of Arabidopsis thaliana. Biochim Biophys Acta. 2009;1787:1230–1237. PubMed

Allahverdiyeva Y, Suorsa M, Rossi F, Pavesi A, Kater MM, Antonacci A, Tadini L, Pribil M, Schneider A, Wanner G, Leister D, Aro EM, Barbato R, Pesaresi P. Arabidopsis plants lacking PsbQ and PsbR subunits of the oxygen-evolving complex show altered PSII super-complex organization and short-term adaptive mechanisms. Plant J. 2013;75:671–684. PubMed

Allen JF, Bennett J, Steinback KE, Arntzen CJ. Chloroplast protein phosphorylation couples plastoquinone redox state to distribution of excitation energy between photosystems. Nature. 1981;291:25–29.

Aro EM, Suorsa M, Rokka A, Allahverdiyeva Y, Paakkarinen V, Saleem A, Battchikova N, Rintamäki E. Dynamics of photosystem II: a proteomic approach to thylakoid protein complexes. J Exp Bot. 2005;56:347–356. PubMed

Bellafiore S, Barneche F, Peltier G, Rochaix JD. State transitions and light adaptation require chloroplast thylakoid protein kinase STN7. Nature. 2005;433:892–895. PubMed

Bennett J. Phosphorylation of chloroplast membrane polypeptides. Nature. 1977;269:344–346.

Ben-Shem A, Frolow F, Nelson N. Crystal structure of plant photosystem I. Nature. 2003;426:630–635. PubMed

Benson SL, Maheswaran P, Ware MA, Hunter CN, Horton P, Jansson S, Ruban AV, Johnson MP. An intact light harvesting complex I antenna system is required for complete state transitions in Arabidopsis. Nat Plants. 2015;1:15176. PubMed

Boekema EJ, van Roon H, Calkoen F, Bassi R, Dekker JP. Multiple types of association of photosystem II and its light-harvesting antenna in partially solubilized photosystem II membranes. Biochemistry. 1999;38:2233–2239. PubMed

Boekema EJ, Jensen PE, Schlodder E, van Breemen JF, van Roon H, Scheller HV, Dekker JP. Green plant photosystem I binds light-harvesting complex I on one side of the complex. Biochemistry. 2001;40:1029–1036. PubMed

Bonaventura C, Myers J. Fluorescence and oxygen evolution from Chlorella pyrenoidosa. Biochim Biophys Acta. 1969;189:366–383. PubMed

Bricker TM, Roose JL, Fagerlund RD, Frankel LK, Eaton-Rye JJ. The extrinsic proteins of Photosystem II. Biochim Biophys Acta. 2012;1817:121–142. PubMed

Buchanan BB, Balmer Y. Redox regulation: a broadening horizon. Annu Rev Plant Biol. 2005;56:187–220. PubMed

Cox J, Mann M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol. 2008;26:1367–1372. PubMed

Crepin A, Caffarri S. The specific localizations of phosphorylated Lhcb1 and Lhcb2 isoforms reveal the role of Lhcb2 in the formation of the PSI–LHCII supercomplex in Arabidopsis during state transitions. Biochim Biophys Acta. 2015;1847:1539–1548. PubMed

Danielsson R, Suorsa M, Paakkarinen V, Albertsson PA, Styring S, Aro EM, Mamedov F. Dimeric and monomeric organization of photosystem II. Distribution of five distinct complexes in the different domains of the thylakoid membrane. J Biol Chem. 2006;281:14241–14249. PubMed

De Las Rivas J, Balsera M, Barber J. Evolution of oxygenic photosynthesis: genome-wide analysis of the OEC extrinsic proteins. Trends Plant Sci. 2004;9:18–25. PubMed

Dekker JP, Boekema EJ. Supramolecular organization of thylakoid membrane proteins in green plants. Biochim Biophys Acta. 2005;1706:12–39. PubMed

Depège N, Bellafiore S, Rochaix J. Role of chloroplast protein kinase Stt7 in LHCII phosphorylation and state transition in Chlamydomonas. Science. 2003;299:1572–1575. PubMed

Derks A, Schaven K, Bruce D. Diverse mechanisms for photoprotection in photosynthesis. Dynamicregulation of photosystem II excitation in response to rapidenvironmental change. Biocim Biophys Acta. 2015;1847:468–485. PubMed

Galka P, Santabarbara S, Khuong TT, Degand H, Morsomme P, Jennings RC, Boekema EJ, Caffarri S. Functional analyses of the plant photosystem I-light-harvesting complex II supercomplex reveal that light-harvesting complex II loosely bound to photosystem II is a very efficient antenna for photosystem I in state II. Plant Cell. 2012;24:2963–2978. PubMed PMC

Grieco M, Tikkanen M, Paakkarinen V, Kangasjärvi S, Aro EM. Steady-state phosphorylation of light-harvesting complex II proteins preserves photosystem I under fluctuating white light. Plant Physiol. 2012;160:1896–1910. PubMed PMC

Grieco M, Suorsa M, Jajoo A, Tikkanen M, Aro EM. Light-harvesting II antenna trimers connect energetically the entire photosynthetic machinery-including both photosystems II and I. Biochim Biophys Acta. 2015;1847:607–619. PubMed

Jansson S. A guide to the Lhc genes and their relatives in Arabidopsis. Trends Plant Sci. 1999;4:236–240. PubMed

Järvi S, Suorsa M, Paakkarinen V, Aro EM. Optimized native gel systems for separation of thylakoid protein complexes: novel super- and mega-complexes. Biochemistry J. 2011;439:207–214. PubMed

Kakiuchi S, Uno C, Ido K, Nishimura T, Noguchi T, Ifuku K, Sato F. The PsbQ protein stabilizes the functional binding of the PsbP protein to photosystem II in higher plants. Biochim Biophys Acta. 2012;1817:1346–1351. PubMed

Kavelaki K, Ghanotakis DF. Effect of the manganese complex on the binding of the extrinsic proteins (17, 23 and 33 kDa) of Photosystem II. Photosynth Res. 1991;29:149–155. PubMed

Koochak H, Puthiyaveetil S, Mullendore DL, Li M, Kirchhoff H. The structural and functional domains of plant thylakoid membranes. Plant J. 2019;97:412–429. PubMed

Koskela MM, Brünje A, Ivanauskaite A, Grabsztunowicz M, Lassowskat I, Neumann U, Dinh TV, Sindlinger J, Schwarzer D, Wirtz M, Tyystjärvi E, Finkemeier I, Mulo P. Chloroplast acetyltransferase NSI is required for state transitions in Arabidopsis thaliana. Plant Cell. 2018;30:1695–1709. PubMed PMC

Kouril R, Zygadlo A, Arteni AA, de Wit CD, Dekker JP, Jensen PE, Scheller HV, Boekema EJ. Structural characterization of a complex of photosystem I and light-harvesting complex II of Arabidopsis thaliana. Biochemistry. 2005;44:10935–10940. PubMed

Kouril R, Dekker JP, Boekema EJ. Supramolecular organization of photosystem II in green plants. Biochim Biophys Acta. 2012;1817:2–12. PubMed

Kyle DJ, Staehelin LA, Arntzen CJ. Lateral mobility of the light-harvesting complex in chloroplast membranes controls excitation energy distribution in higher plants. Arch Biochem Biophys. 1983;222:527–541. PubMed

Li Y, Liu B, Zhang J, Kong F, Zhang L, Meng H, Li W, Rochaix JD, Li D, Peng L. OHP1, OHP2, and HCF244 form a transient functional complex with the Photosystem II reaction center. Plant Physiol. 2019;179:195–208. PubMed PMC

Lohscheider JN, Rojas-Stutz MC, Rothbart M, Andersson U, Funck D, Mendgen K, Grimm B, Adamska I. Altered levels of LIL3 isoforms in Arabidopsis lead to disturbed pigment-protein assembly and chlorophyll synthesis, chlorotic phenotype and impaired photosynthetic performance. Plant Cell Environ. 2015;38:2115–2127. PubMed

Longoni P, Douchi D, Cariti F, Fucile G, Goldschmidt-Clermont M. Phosphorylation of the light-harvesting complex II isoform Lhcb2 is central to state transitions. Plant Physiol. 2015;169:2874–2883. PubMed PMC

Lunde C, Jensen PE, Haldrup A, Knoetzel J, Scheller HV. The PSI-H subunit of photosystem I is essential for state transitions in plant photosynthesis. Nature. 2000;408:613–615. PubMed

Miyao M, Murata N. Effect of urea on photosystem II particles: evidence for an essential role of the 33 kilodalton polypeptide in photosynthetic oxygen evolution. Biochim Biophys Acta. 1984;765:253–257.

Morgan MJ, Lehmann M, Schwarzlander M, Baxter CJ, Sienkiewicz-Porzucek A, Williams TC, Schauer N, Fernie AR, Fricker MD, Ratcliffe RG, Sweetlove LJ, Finkemeier I. Decrease in manganese superoxide dismutase leads to reduced root growth and affects tricarboxylic acid cycle flux and mitochondrial redox homeostasis. Plant Physiol. 2008;147:101–114. PubMed PMC

Murata N. Control of excitation transfer in photosynthesis. I. Light-induced change of chlorophyll a fluorescence in Porphyridium cruentum. Biochim Biophys Acta. 1969;172:242–251. PubMed

Myouga F, Takahashi K, Tanaka R, Nagata N, Kiss AZ, Funk C, Nomura Y, Nakagami H, Jansson S, Shinozaki K. Stable accumulation of Photosystem II requires One-Helix Protein1 (OHP1) of the light harvesting-like family. Plant Physiol. 2018;176:2277–2291. PubMed PMC

Pan X, Ma J, Su X, Cao P, Chang W, Liu Z, Zhang X, Li M. Structure of the maize photosystem I supercomplex with light-harvesting complexes I and II. Science. 2018;360:1109–1113. PubMed

Pesaresi P, Hertle A, Pribil M, Kleine T, Wagner R, Strissel H, Ihnatowicz A, Bonardi V, Scharfenberg M, Schneider A, Pfannschmidt T, Leister D. Arabidopsis STN7 kinase provides a link between short- and long-term photosynthetic acclimation. Plant Cell. 2009;21:2402–2423. PubMed PMC

Pietrzykowska M, Suorsa M, Semchonok DA, Tikkanen M, Boekema EJ, Aro EM, Jansson S. The light-harvesting chlorophyll a/b binding proteins Lhcb1 and Lhcb2 play complementary roles during state transitions in Arabidopsis. Plant Cell. 2014;26:3646–3660. PubMed PMC

Porra RJ, Thompson WA, Kriedemann PE. Determination of accurate extinction coefficients and simultaneous-equations for assaying chlorophyll-A and chlorophyll-B extracted with 4 different solvents-verification of the concentration of chlorophyll standards by atomic-absorption spectroscopy. Biochim Biophys Acta. 1989;975:384–394.

Pribil M, Pesaresi P, Hertle A, Barbato R, Leister D. Role of plastid protein phosphatase TAP38 in LHCII dephosphorylation and thylakoid electron flow. PLoS Biol. 2010;8:e1000288. PubMed PMC

Pribil M, Labs M, Leister D. Structure and dynamics of thylakoids in land plants. J Exp Bot. 2014;65:1955–1972. PubMed

Puthiyaveetil S, Ibrahim IM, Jelicic B, Tomasic A, Fulgosi H, Allen JF. Transcriptional control of photosynthesis genes: the evolutionarily conserved regulatory mechanism in plastid genome function. Genome Biol Evol. 2010;2:888–896. PubMed PMC

Puthiyaveetil S, Ibrahim IM, Allen JF. Oxidation-reduction signalling components in regulatory pathways of state transitions and photosystem stoichiometry adjustment in chloroplasts. Plant Cell Environ. 2012;35:347–359. PubMed

Rantala M, Tikkanen M, Aro EM. Proteomic characterization of hierarchical megacomplex formation in Arabidopsis thylakoid membrane. Plant J. 2017;92:951–962. PubMed

Rochaix JD. Regulation and dynamics of the light-harvesting system. Annu Rev Plant Biol. 2014;65:287–309. PubMed

Rochaix JD, Bassi R. LHC-like proteins involved in stress responses and biogenesis/repair of the photosynthetic apparatus. Biochem J. 2019;476:581–593. PubMed

Schneider D, Lopez LS, Li M, Crawford JD, Kirchhoff H, Kunz H-H. Fluctuating light experiments and semi-automated plant phenotyping enabled by self-built growth racks and simple upgrades to the IMAGING-PAM. Plant Methods. 2019;15:156. PubMed PMC

Schöttler MA, Albus CA, Bock R. Photosystem I: its biogenesis and function in higher plants. J Plant Physiol. 2011;168:1452–1461. PubMed

Schwanhäusser B, Busse D, Li N, Dittmar G, Schuchhardt J, Wolf J, Chen W, Selbach M. Global quantification of mammalian gene expression control. Nature. 2011;473:337–342. PubMed

Shapiguzov A, Ingelsson B, Samol I, Andres C, Kessler F, Rochaix JD, Vener AV, Goldschmidt-Clermont M. The PPH1 phosphatase is specifically involved in LHCII dephosphorylation and state transitions in Arabidopsis. Proc Natl Acad Sci USA. 2010;107:4782–4787. PubMed PMC

Steiner S, Dietzel L, Schröter Y, Fey V, Wagner R, Pfannschmidt T. The role of phosphorylation in redox regulation of photosynthesis genes psaA and psbA during photosynthetic acclimation of mustard. Mol Plant. 2009;2:416–429. PubMed

Suorsa M, Rantala M, Danielsson R, Järvi S, Paakkarinen V, Schröder WP, Styring S, Mamedov F, Aro EM. Dark-adapted spinach thylakoid protein heterogeneity offers insights into the photosystem II repair cycle. Biochim Biophys Acta. 2014;1837:1463–1471. PubMed

Suorsa M, Rantala M, Mamedov F, Lespinasse M, Trotta A, Grieco M, Vuorio E, Tikkanen M, Järvi S, Aro EM. Light acclimation involves dynamic re-organization of the pigment-protein megacomplexes in non-appressed thylakoid domains. Plant J. 2015;84:360–373. PubMed

Tanaka R, Rothbart M, Oka S, Takabayashi A, Takahashi K, Shibata M, Myouga F, Motohashi R, Shinozaki K, Grimm B, Tanaka A. LIL3, a light-harvesting-like protein, plays an essential role in chlorophyll and tocopherol biosynthesis. Proc Natl Acad Sci USA. 2010;107:16721–16725. PubMed PMC

Tikkanen M, Piippo M, Suorsa M, Sirpiö S, Mulo P, Vainonen J, Vener AV, Allahverdiyeva Y, Aro EM. State transitions revisited-a buffering system for dynamic low light acclimation of Arabidopsis. Plant Mol Biol. 2006;62:779–793. PubMed

Tikkanen M, Nurmi M, Kangasjärvi S, Aro EM. Core protein phosphorylation facilitates the repair of photodamaged photosystem II at high light. Biochim Biophys Acta. 2008;1777:1432–1437. PubMed

Tikkanen M, Grieco M, Kangasjärvi S, Aro EM. Thylakoid protein phosphorylation in higher plant chloroplasts optimizes electron transfer under fluctuating light. Plant Physiol. 2010;152:723–735. PubMed PMC

Tikkanen M, Suorsa M, Gollan PJ, Aro EM. Post-genomic insight into thylakoid membrane lateral heterogeneity and redox balance. FEBS Lett. 2012;586:2911–2916. PubMed

Trotta A, Bajwa AA, Mancini I, Paakkarinen V, Pribil M, Aro E-M. The role of phosphorylation dynamics of Curvature Thylakoid 1B in plant thylakoid membranes. Plant Phys. 2019;181:1615–1631. PubMed PMC

Tyanova S, Temu T, Sinitcyn P, Carlson A, Hein MY, Geiger T, Mann M, Cox J. The Perseus computational platform for comprehensive analysis of (prote)omics data. Nat Methods. 2016;13:731–740. PubMed

Wientjes E, van Amerongen H, Croce R. LHCII is an antenna of both photosystems after long-term acclimation. Biochim Biophys Acta. 2013;1827:420–426. PubMed