High-light-inducible proteins (Hlips) are single-helix transmembrane proteins that are essential for the survival of cyanobacteria under stress conditions. The model cyanobacterium Synechocystis sp. PCC 6803 contains four Hlip isoforms (HliA-D) that associate with Photosystem II (PSII) during its assembly. HliC and HliD are known to form pigmented (hetero)dimers that associate with the newly synthesized PSII reaction center protein D1 in a configuration that allows thermal dissipation of excitation energy. Thus, it is expected that they photoprotect the early steps of PSII biogenesis. HliA and HliB, on the other hand, bind the PSII inner antenna protein CP47, but the mode of interaction and pigment binding have not been resolved. Here, we isolated His-tagged HliA and HliB from Synechocystis and show that these two very similar Hlips do not interact with each other as anticipated, rather they form HliAC and HliBC heterodimers. Both dimers bind Chl and β-carotene in a quenching conformation and associate with the CP47 assembly module as well as later PSII assembly intermediates containing CP47. In the absence of HliC, the cellular levels of HliA and HliB were reduced, and both bound atypically to HliD. We postulate a model in which HliAC-, HliBC-, and HliDC-dimers are the functional Hlip units in Synechocystis. The smallest Hlip, HliC, acts as a 'generalist' that prevents unspecific dimerization of PSII assembly intermediates, while the N-termini of 'specialists' (HliA, B or D) dictate interactions with proteins other than Hlips.

Institute of Chemistry Faculty of Science University of South Bohemia Branišovská 1760 37005 České Budějovice Czech Republic

Institute of Microbiology of the Czech Academy of Sciences Novohradská 237 Opatovický mlýn 37901 Třeboň Czech Republic

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Photosynth Res. 2023 Jul;157(1):53. doi: 10.1007/s11120-023-01016-y. PubMed

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Arkowitz RA, Wickner W (1994) SecD and SecF are required for the proton electrochemical gradient stimulation of preprotein translocation. EMBO J 13:954–963. https://doi.org/10.1002/j.1460-2075.1994.tb06340.x PubMed DOI PMC

Bassi R, Croce R, Cugini D, Sandona D (1999) Mutational analysis of a higher plant antenna protein provides identification of chromophores bound into multiple sites. Proc Natl Acad Sci 96:10056–10061. https://doi.org/10.1073/pnas.96.18.10056 PubMed DOI PMC

Becker K, Cormann KU, Nowaczyk MM (2011) Assembly of the water-oxidizing complex in photosystem II. J Photochem Photobiol B 104:204–211. https://doi.org/10.1016/j.jphotobiol.2011.02.005 PubMed DOI

Ben-Shem A, Frolow F, Nelson N (2003) Crystal structure of plant photosystem I. Nature 426:630–635. https://doi.org/10.1038/nature02200 PubMed DOI

Boehm M, Romero E, Reisinger V, Yu J, Komenda J, Eichacker LA, Dekker JP, Nixon PJ (2011) Investigating the early stages of photosystem II assembly in Synechocystis sp. PCC 6803. J Biol Chem 286:14812–14819. https://doi.org/10.1074/jbc.M110.207944 PubMed DOI PMC

Boehm M, Yu J, Reisinger V, Bečková M, Eichacker LA, Schlodder E, Komenda J, Nixon PJ (2012) Subunit composition of CP43-less photosystem II complexes of Synechocystis sp. PCC 6803: implications for the assembly and repair of photosystem II. Philos Trans R Soc B 367:3444–3454. https://doi.org/10.1098/rstb.2012.0066 DOI

Botte M, Zaccai NR, Lycklama à J, Nijeholt JL, Martin R, Knoops K, Papai G, Zou J, Deniaud A, Karuppasamy M, Jiang Q, Roy AS, Schulten K, Schultz P, Rappsilber J, Zaccai G, Berger I, Collinson I, Schaffitzel C (2016) A central cavity within the holo-translocon suggests a mechanism for membrane protein insertion. Sci Rep 6:38399. https://doi.org/10.1038/srep38399

Chae PS, Rasmussen SGF, Rana RR, Gotfryd K, Kruse AC, Manglik A, Cho KH, Nurva S, Gether U, Guan L, Loland CJ, Byrne B, Kobilka BK, Gellman SH (2012) A new class of amphiphiles bearing rigid hydrophobic groups for solubilization and stabilization of membrane proteins. Chem Eur J 18:9485–9490. https://doi.org/10.1002/chem.201200069 PubMed DOI

Chidgey J, Linhartová M, Komenda J, Jackson PJ, Dickman MJ, Canniffe DP, Koník P, Pilný J, Hunter CN, Sobotka R (2014) A cyanobacterial chlorophyll synthase-HliD complex associates with the Ycf39 protein and the YidC/Alb3 insertase. Plant Cell 26:1267–1279. https://doi.org/10.1105/tpc.114.124495 PubMed DOI PMC

Cox J, Mann M (2008) MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol 26(12):1367–1372. https://doi.org/10.1038/NBT.1511 PubMed DOI

Cox J, Neuhauser N, Michalski A, Scheltema RA, Olsen JV, Mann M (2011) Andromeda: a peptide search engine integrated into the MaxQuant environment. J Proteome Res 10:1794–1805. https://doi.org/10.1021/pr101065j PubMed DOI

Dobáková M, Sobotka R, Tichý M, Komenda J (2009) Psb28 protein is involved in the biogenesis of the photosystem II inner antenna CP47 (PsbB) in the cyanobacterium Synechocystis sp. PCC 6803. Plant Physiol 149:1076–1086. https://doi.org/10.1104/pp.108.130039 PubMed DOI PMC

Dolganov NA, Bhayat D, Grossman AR (1995) Cyanobacterial protein with similarity to the chlorophyll a/b binding proteins of higher plants: Evolution and regulation. Proc Natl Acad Sci USA 92:636–640. https://doi.org/10.1073/pnas.92.2.636 PubMed DOI PMC

Engelken J, Brinkmann H, Adamska I (2010) Taxonomic distribution and origins of the extended LHC (light-harvesting complex) antenna protein superfamily. BMC Evol Biol 10:233. https://doi.org/10.1186/1471-2148-10-233 PubMed DOI PMC

Gardel C, Johnson K, Jacq A, Beckwith J (1990) The secD locus of E. coli codes for two membrane proteins required for protein export. EMBO J 9:3209–3216. https://doi.org/10.1002/j.1460-2075.1990.tb07519.x PubMed DOI PMC

Havaux M, Guedeney G, He Q, Grossman AR (2003) Elimination of high-light-inducible polypeptides related to eukaryotic chlorophyll a/b-binding proteins results in aberrant photoacclimation in Synechocystis PCC6803. Biochim Biophys Acta Bioenerget 1557:21–33. https://doi.org/10.1016/S0005-2728(02)00391-2 DOI

He Q, Dolganov N, Björkman O, Grossman AR (2001) The high light-inducible polypeptides in Synechocystis PCC6803. Expression and function in high light. J Biol Chem 276:306–314. https://doi.org/10.1074/jbc.M008686200 PubMed DOI

Hey D, Grimm B (2018) ONE-HELIX PROTEIN2 (OHP2) is required for the stability of OHP1 and assembly factor HCF244 and is functionally linked to PSII biogenesis. Plant Physiol 177:1453–1472. https://doi.org/10.1104/pp.18.00540 PubMed DOI PMC

Hey D, Grimm B (2020) ONE-HELIX PROTEIN1 and 2 form heterodimers to bind chlorophyll in photosystem II biogenesis. Plant Physiol 183:179–193. https://doi.org/10.1104/pp.19.01304 PubMed DOI PMC

Hollingshead S, Kopečná J, Jackson PJ, Canniffe DP, Davison PA, Dickman MJ, Sobotka R, Hunter CN (2012) Conserved chloroplast open-reading frame ycf54 is required for activity of the magnesium protoporphyrin monomethylester oxidative cyclase in Synechocystis PCC 6803. J Biol Chem 287:27823–27833. https://doi.org/10.1074/jbc.M112.352526

Hontani Y, Kloz M, Polívka T, Shukla MK, Sobotka R, Kennis JTM (2018) Molecular origin of photoprotection in cyanobacteria probed by watermarked femtosecond stimulated Raman spectroscopy. J Phys Chem Lett 9:1788–1792. https://doi.org/10.1021/acs.jpclett.8b00663 PubMed DOI PMC

Järvi S, Suorsa M, Aro EM (2015) Photosystem II repair in plant chloroplasts—regulation, assisting proteins and shared components with photosystem II biogenesis. Biochim Biophys Acta (BBA) - Bioenerg 1847:900–909. https://doi.org/10.1016/J.BBABIO.2015.01.006 DOI

Jordan P, Fromme P, Witt HT, Klukas O, Saenger W, Krauß N (2001) Three-dimensional structure of cyanobacterial photosystem I at 2.5 Å resolution. Nature 411:909–917. https://doi.org/10.1038/35082000 PubMed DOI

Kameo S, Aso M, Furukawa R, Matsumae R, Yokono M, Fujita T, Tanaka A, Tanaka R, Takabayashi A (2021) Substitution of deoxycholate with the amphiphilic polymer amphipol A8–35 improves the stability of large protein complexes during native electrophoresis. Plant Cell Physiol 62:348–355. https://doi.org/10.1093/pcp/pcaa165 PubMed DOI

Knoppová J, Sobotka R, Tichý M, Yu J, Konik P, Halada P, Nixon PJ, Komenda J (2014) Discovery of a chlorophyll binding protein complex involved in the early steps of photosystem II assembly in Synechocystis. Plant Cell 26:1200–1212. https://doi.org/10.1105/tpc.114.123919 PubMed DOI PMC

Knoppová J, Yu J, Konik P, Nixon PJ, Komenda J (2016) CyanoP is involved in the early steps of photosystem II assembly in the cyanobacterium Synechocystis sp. PCC 6803. Plant Cell Physiol 57:1921–1931. https://doi.org/10.1093/pcp/pcw115 PubMed DOI

Komar J, Alvira S, Schulze RJ, Martin R, Nijeholt JL, Lee SC, Dafforn TR, Deckers-Hebestreit G, Berger I, Schaffitzel C, Collinson I (2016) Membrane protein insertion and assembly by the bacterial holo-translocon SecYEG–SecDF–YajC–YidC. Biochem J 473:3341–3354. https://doi.org/10.1042/BCJ20160545

Komenda J, Sobotka R (2016) Cyanobacterial high-light-inducible proteins - Protectors of chlorophyll-protein synthesis and assembly. Biochim Biophys Acta - Bioenerg 1857:288–295. https://doi.org/10.1016/j.bbabio.2015.08.011 DOI

Komenda J, Nickelsen J, Tichý M, Prášil O, Eichacker LA, Nixon PJ (2008) The cyanobacterial homologue of HCF136/YCF48 is a component of an early photosystem II assembly complex and is important for both the efficient assembly and repair of photosystem II in Synechocystis sp. PCC 6803. J Biol Chem 283:22390–22399. https://doi.org/10.1074/jbc.M801917200 PubMed DOI

Komenda J, Knoppová J, Kopečná J, Sobotka R, Halada P, Yu J, Nickelsen J, Boehm M, Nixon PJ (2012a) The Psb27 assembly factor binds to the CP43 complex of photosystem II in the cyanobacterium Synechocystis sp. PCC 6803. Plant Physiol 158:476–486. https://doi.org/10.1104/pp.111.184184 PubMed DOI

Komenda J, Sobotka R, Nixon PJ (2012b) Assembling and maintaining the photosystem II complex in chloroplasts and cyanobacteria. Curr Opin Plant Biol 15:245–251. https://doi.org/10.1016/j.pbi.2012.01.017 PubMed DOI

Komenda J, Krynická V, Zakar T (2019) Isolation of thylakoid membranes from the cyanobacterium Synechocystis sp. PCC 6803 and analysis of their photosynthetic pigment-protein complexes by clear native-PAGE. Bio-Protoc 9:3126. https://doi.org/10.21769/BIOPROTOC.3126 DOI

Kühlbrandt W, Wang DN, Fujiyoshi Y (1994) Atomic model of plant light-harvesting complex by electron crystallography. Nature 367:614–621. https://doi.org/10.1038/367614a0 PubMed DOI

Lee J, Lee HJ, Shin MK, Ryu WS (2004) Versatile PCR-mediated insertion or deletion mutagenesis. Biotechniques 36:398–400. https://doi.org/10.2144/04363BM04 PubMed DOI

Lichtenthaler HK, Wellburn AR (1983) Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592 DOI

Liu Z, Yan H, Wang K, Kuang T, Gui L, An X, Chang W (2004) Crystal structure of spinach major light-harvesting complex at 2.72 Å resolution. Nature 428:287–292. https://doi.org/10.1038/nature02373 PubMed DOI

Li Y, Liu B, Zhang J, Kong F, Meng H, Li W, Rochaix JD, Li D, Peng L (2019) OHP1, OHP2, and HCF244 form a transient functional complex with the photosystem II reaction center. Plant Physiol 179:195–208. https://doi.org/10.1104/pp.18.01231 PubMed DOI

Llansola-Portoles MJ, Sobotka R, Kish E, Shukla MK, Pascal AA, Polívka T, Robert B (2017) Twisting a β-carotene, an adaptive trick from nature for dissipating energy during photoprotection. J Biol Chem 292:1396. https://doi.org/10.1074/JBC.M116.753723 PubMed DOI

Madeira F, Park YM, Lee J, Buso N, Gur T, Madhusoodanan N, Basutkar P, Tivey ARN, Potter SC, Finn RD, Lopez R (2019) The EMBL-EBI search and sequence analysis tools APIs in 2019. Nucleic Acids Res 47:W636–W641. https://doi.org/10.1093/nar/gkz268 PubMed DOI PMC

Malavath T, Caspy I, Netzer-El SY, Klaiman D, Nelson N (2018) Structure and function of wild-type and subunit-depleted photosystem I in Synechocystis. Biochim Biophys Acta - Bioenerg 1859:645–654. https://doi.org/10.1016/j.bbabio.2018.02.002 PubMed DOI

Mazor Y, Borovikova A, Caspy I, Nelson N (2017) Structure of the plant photosystem I supercomplex at 2.6 Å resolution. Nat Plants 3:17014. https://doi.org/10.1038/nplants.2017.14 PubMed DOI

Myouga F, Takahashi K, Tanaka R, Nagata N, Kiss AS, Funk C, Nomura Y, Nakagami H, Jansson S, Shinozaki K (2018) Stable accumulation of photosystem II requires ONE-HELIX PROTEIN1 (OHP1) of the light harvesting-like family. Plant Physiol 176:2277–2291. https://doi.org/10.1104/pp.17.01782 PubMed DOI PMC

Niedzwiedzki DM, Tronina T, Liu H, Staleva H, Komenda J, Sobotka R, Blankenship RE, Polívka T (2016) Carotenoid-induced non-photochemical quenching in the cyanobacterial chlorophyll synthase-HliC/D complex. Biochim Biophys Acta - Bioenerg 1857:1430–1439. https://doi.org/10.1016/j.bbabio.2016.04.280 DOI

Pascual-Aznar G, Konert G, Bečková M, Kotabová E, Gardian Z, Knoppová J, Bučinská L, Kaňa T, Sobotka R, Komenda J (2021) Psb35 protein stabilizes the CP47 assembly module and associated high-light inducible proteins during the biogenesis of photosystem ii in the cyanobacterium Synechocystis sp. PCC6803. Plant Cell Physiol 62:178–190. https://doi.org/10.1093/pcp/pcaa148 PubMed DOI

Pazderník M, Mareš J, Pilný J, Sobotka R (2019) The antenna-like domain of the cyanobacterial ferrochelatase can bind chlorophyll and carotenoids in an energy-dissipative configuration. J Biol Chem 294:11131–11143. https://doi.org/10.1074/jbc.ra119.008434 PubMed DOI PMC

Pogliano JA, Beckwith J (1994) SecD and SecF facilitate protein export in Escherichia coli. EMBO J 13:554–561. https://doi.org/10.1002/j.1460-2075.1994.tb06293.x PubMed DOI PMC

Proctor MS, Pazderník M, Jackson PJ, Pilný J, Martin EC, Dickman MJ, Canniffe DP, Johnson MP, Hunter CN, Sobotka R, Hitckcock A (2020) Xanthophyll carotenoids stabilise the association of cyanobacterial chlorophyll synthase with the LHC-like protein HliD. Biochem J 477:4021–4036. https://doi.org/10.1042/BCJ20200561 PubMed DOI

Promnares K, Komenda J, Bumba L, Nebesarova J, Vácha F, Tichý M (2006) Cyanobacterial small chlorophyll-binding protein ScpD (HliB) is located on the periphery of photosystem II in the vicinity of PsbH and CP47 subunits. J Biol Chem 281:32705–32713. https://doi.org/10.1074/jbc.M606360200 PubMed DOI

Rahimzadeh Karvansara P, Pascual Aznar G, Bečková M, Komenda J (2022) The Psb34 protein modulates binding of high-light-inducible proteins to CP47 containing photosystem II assembly intermediates in the cyanobacterium Synechocystis sp. PCC 6803. Photosynth Res, accepted

Rappsilber J, Mann M, Ishihama Y (2007) Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nat Protoc 2(8):1896–1906. https://doi.org/10.1038/NPROT.2007.261 PubMed DOI

Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W, Smyth GK (2015) Limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res 43:e47. https://doi.org/10.1093/nar/gkv007 PubMed DOI PMC

Rudolph JD, Cox J (2019) A network module for the Perseus software for computational proteomics facilitates proteome interaction graph analysis. J Proteome Res 18:2052–2064. https://doi.org/10.1021/acs.jproteome.8b00927 PubMed DOI PMC

Shukla MK, Llansola-Portoles MJ, Tichý M, Pascal AA, Robert B, Sobotka R (2018) Binding of pigments to the cyanobacterial high-light-inducible protein HliC. Photosynth Res 137:29–39. https://doi.org/10.1007/s11120-017-0475-7 PubMed DOI

Skotnicová P, Staleva H, Kuznetsova V, Bína D, Konert M, Lu S, Polivka T, Sobotka R (2021) Plant LHC-like proteins show robust folding and static non-photochemical quenching. Nat Commun 12:6890. https://doi.org/10.1038/s41467-021-27155-1 PubMed DOI PMC

Staleva H, Komenda J, Shukla MK, Šlouf V, Kaňa R, Polívka T, Sobotka R (2015) Mechanism of photoprotection in the cyanobacterial ancestor of plant antenna proteins. Nat Chem Biol 11:287–291. https://doi.org/10.1038/nchembio.1755 PubMed DOI

Tichý M, Bečková M, Kopečná J, Noda J, Sobotka R, Komenda J (2016) Strain of Synechocystis PCC 6803 with aberrant assembly of photosystem II contains tandem duplication of a large chromosomal region. Front Plant Sci 7:648. https://doi.org/10.3389/FPLS.2016.00648 PubMed DOI PMC

Tribet C, Audebert R, Popot J-L (1996) Amphipols: polymers that keep membrane proteins soluble in aqueous solutions. Proc Natl Acad Sci 93:15047–15050. https://doi.org/10.1073/pnas.93.26.15047 PubMed DOI PMC

Tyanova S, Temu T, Cox J (2016a) The MaxQuant computational platform for mass spectrometry-based shotgun proteomics. Nat Protoc 11:2301–2319. https://doi.org/10.1038/NPROT.2016.136 PubMed DOI

Tyanova S, Temu T, Sinitcyn P, Carlson A, Hein MY, Geiger T, Mann M, Cox J (2016b) The Perseus computational platform for comprehensive analysis of (prote)omics data. Nat Methods 13:731–740. https://doi.org/10.1038/nmeth.3901 PubMed DOI

Umena Y, Kawakami K, Shen JR, Kamiya N (2011) Crystal structure of oxygen-evolving photosystem II at a resolution of 1.9Å. Nature 473:55–60. https://doi.org/10.1038/nature09913 PubMed DOI

Vass I (2012) Molecular mechanisms of photodamage in the photosystem II complex. Biochim Biophys Acta (BBA) - Bioenerg 1817:209–217. https://doi.org/10.1016/J.BBABIO.2011.04.014 DOI

Wei X, Su X, Cao P, Liu X, Chang W, Li M, Zhang X, Liu Z (2016) Structure of spinach photosystem II-LHCII supercomplex at 3.2 Å resolution. Nature 534:69–74. https://doi.org/10.1038/nature18020 PubMed DOI

Xiao Y, Huang G, You X, Wang W, Kuang T, Han G, Sui SF, Shen JR (2021) Structural insights into cyanobacterial photosystem II intermediates associated with Psb28 and Tsl0063. Nat Plants 7:1132–1142. https://doi.org/10.1038/s41477-021-00961-7 PubMed DOI

Xu H, Vavilin D, Funk C, Vermaas WFJ (2002) Small Cab-like proteins regulating tetrapyrrole biosynthesis in the cyanobacterium Synechocystis sp. PCC 6803 Plant Mol Biol 49:149–160. https://doi.org/10.1023/A:1014900806905

Yao D, Kieselbach T, Komenda J, Promnares K, Hernández Prieto MA, Tichy M, Vermaas W, Funk C (2007) Localization of the small CAB-like proteins in photosystem II. J Biol Chem 282:267–276. https://doi.org/10.1074/jbc.M605463200 PubMed DOI

Zabret J, Bohn S, Schuller S, Arnolds O, Möller M, Meier-Credo J, Liauw P, Chan A, Tajkhorshid E, Langer J, Stoll R, Krieger-Liszkay A, Engel B, Rudack T, Schuller J, Nowaczyk M (2021) Structural insights into photosystem II assembly. Nat Plants 7:524–538. https://doi.org/10.1038/s41477-021-00895-0 PubMed DOI PMC

The biogenesis and maintenance of PSII: Recent advances and current challenges

FtsH4 protease controls biogenesis of the PSII complex by dual regulation of high light-inducible proteins

Psb34 protein modulates binding of high-light-inducible proteins to CP47-containing photosystem II assembly intermediates in the cyanobacterium Synechocystis sp. PCC 6803