In the model cyanobacterium Synechocystis sp. PCC 6803, the terminal enzyme of chlorophyll biosynthesis, chlorophyll synthase (ChlG), forms a complex with high light-inducible proteins, the photosystem II assembly factor Ycf39 and the YidC/Alb3/OxaI membrane insertase, co-ordinating chlorophyll delivery with cotranslational insertion of nascent photosystem polypeptides into the membrane. To gain insight into the ubiquity of this assembly complex in higher photosynthetic organisms, we produced functional foreign chlorophyll synthases in a cyanobacterial host. Synthesis of algal and plant chlorophyll synthases allowed deletion of the otherwise essential native cyanobacterial gene. Analysis of purified protein complexes shows that the interaction with YidC is maintained for both eukaryotic enzymes, indicating that a ChlG-YidC/Alb3 complex may be evolutionarily conserved in algae and plants.

Department of Chemical and Biological Engineering ChELSI Institute University of Sheffield UK

Department of Molecular Biology and Biotechnology University of Sheffield UK

Faculty of Science University of South Bohemia České Budějovice Czech Republic

Institute of Microbiology Czech Academy of Sciences Center Algatech Třeboň Czech Republic

See more in PubMed

Jordan P, Fromme P, Witt HT, Klukas O, Saenger W and Krausz N (2001) Three‐dimensional structure of cyanobacterial photosystem I at 2.5 Å resolution. Nature 411, 909–917. PubMed

Umena Y, Kawakami K, Shen J‐R and Kamiya N (2011) Crystal structure of oxygen‐evolving photosystem II at a resolution of 1.9Å. Nature 473, 55–60. PubMed

Wei X, Su X, Cao P, Liu X, Chang W, Li M, Zhang X and Liu Z (2016) Structure of spinach photosystem II‐LHCII supercomplex at 3.2 Å resolution. Nature 534, 69–74. PubMed

Mazor Y, Borovikova A, Caspy I and Nelson N (2017) Structure of the plant photosystem I supercomplex at 2.6 Å resolution. Nat Plants 3, 17014. PubMed

Nickelsen J and Rengstl B (2013) Photosystem II assembly: from cyanobacteria to plants. Annu Rev Plant Biol 64, 609–635. PubMed

Yang H, Liu J, Wen X and Lu C (2015) Molecular mechanism of photosystem I assembly in oxygenic organisms. Biochim Biophys Acta 1847, 838–848. PubMed

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

Komenda J and Sobotka R (2016) Cyanobacterial high‐light‐inducible proteins ‐ Protectors of chlorophyll‐protein synthesis and assembly. Biochim Biophys Acta 1857, 288–295. PubMed

Bhaya D, Dufresne A, Vaulot D and Grossman A (2002) Analysis of the hli gene family in marine and freshwater cyanobacteria. FEMS Microbiol Lett 215, 209–219. PubMed

Staleva H, Komenda J, Shukla MK, Šlouf V, Kaňa R, Polívka T and Sobotka R (2015) Mechanism of photoprotection in the cyanobacterial ancestor of plant antenna proteins. Nat Chem Biol 11, 287–291. PubMed

Niedzwiedzki DM, Tronina T, Liu H, Staleva H, Komenda J, Sobotka R, Blankenship RE and Polívka T (2016) Carotenoid‐induced non‐photochemical quenching in the cyanobacterial chlorophyll synthase‐HliC/D complex. Biochim Biophys Acta 1857, 1430–1439. PubMed

Kallberg Y, Oppermann U, Jörnvall H and Persson B (2002) Short‐chain dehydrogenases/reductases (SDRs). Coenzyme‐based functional assignments in completed genomes. Eur J Biochem 269, 4409–4417. PubMed

Knoppová J, Sobotka R, Tichý M, Yu J, Konik P, Halada P, Nixon PJ and 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. PubMed PMC

Beck K, Eisner G, Trescher D, Dalbey RE, Brunner J and Müller M (2001) YidC, an assembly site for polytopic Escherichia coli membrane proteins located in immediate proximity to the SecYE translocon and lipids. EMBO Rep 2, 709–714. PubMed PMC

Nagamori S, Smirnova IN and Kaback HR (2004) Role of YidC in folding of polytopic membrane proteins. J Cell Biol 165, 53–62. PubMed PMC

Spence E, Bailey S, Nenninger A, Møller SG and Robinson C (2004) A homolog of Albino3/OxaI is essential for thylakoid biogenesis in the cyanobacterium Synechocystis sp. PCC6803. J Biol Chem 279, 55792–55800. PubMed

Göhre V, Ossenbühl F, Crèvecoeur M, Eichacker LA and Rochaix J‐D (2006) One of two Alb3 proteins is essential for the assembly of the photosystems and for cell survival in Chlamydomonas . Plant Cell 18, 1454–1466. PubMed PMC

Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R et al (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23, 2947–2948. PubMed

Kearse M, Moir R, Wilson A, Stones‐Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C et al (2012) Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28, 1647–1649. PubMed PMC

Rippka R, Derueles J, Waterbury JB, Herdman M and Stainer RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111, 1–61.

Emanuelsson O, Nielsen H and von Heijne G (1999) ChloroP, a neural network‐based method for predicting chloroplast transit peptides and their cleavage sites. Protein Sci 8, 978–984. PubMed PMC

Hollingshead S, Kopečná J, Jackson PJ, Canniffe DP, Davison PA, Dickman MJ, Sobotka R and 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. PubMed PMC

Largarde D and Vermaas W (1999) The zeaxanthin biosynthesis enzyme beta‐carotene hydroxylase is involved in myxoxanthophyll synthesis in Synechocystis sp. PCC 6803. FEBS Lett 454, 247–251. PubMed

Porra RJ, Thompson WA and Kriedemann PE (1989) 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.

Pilný J, Kopečná J, Noda J and Sobotka R (2015) Detection and quantification of heme and chlorophyll precursors using a high performance liquid chromatography (HPLC) system equipped with two fluorescence detectors. Bio‐Protocol 5, e1390.

Oster U, Bauer CE and Rüdiger W (1997) Characterization of chlorophyll a and bacteriochlorophyll a synthases by heterologous expression in Escherichia coli . J Biol Chem 272, 9671–9676–635. PubMed

Kim J, Klein PG and Mullet JE (1994) Synthesis and turnover of photosystem II reaction center protein D1. Ribosome pausing increases during chloroplast development. J Biol Chem 269, 17918–17923. PubMed

Zhang L, Paakkarinen V, van Wijk KJ and Aro E‐M (1999) Co‐translational assembly of the D1 protein into photosystem II. J Biol Chem 274, 16062–16067. PubMed

Funk C and Vermaas W (1999) A cyanobacterial gene family coding for single‐helix proteins resembling part of the light‐harvesting proteins from higher plants. Biochemistry 38, 9397–9404. PubMed

Xu H, Vavilin D, Funk C and Vermaas W (2004) Multiple deletions of small cab‐like proteins in the cyanobacterium Synechocystis sp. PCC 6803. Consequences for pigment biosynthesis and accumulation. J Biol Chem 279, 27971–27979. PubMed

Sinha RK, Komenda J, Knoppová J, Sedlářová M and Pospíšil P (2012) Small CAB‐like proteins prevent formation of singlet oxygen in the damaged photosystem II complex of the cyanobacterium Synechocystis sp. PCC 6803. Plant, Cell Environ 35, 806–818. PubMed

Beck J, Lohscheider JN, Albert S, Andersson U, Mendgen KW, Rojas‐Stütz MC, Adamska I and Funck D (2017) Small one‐helix proteins are essential for photosynthesis in arabidopsis. Front Plant Sci 8, 1–14. PubMed PMC

Myouga F, Takahashi K, Tanaka R, Nagata N, Kiss AZ, Funk C, Nomura Y, Nakagami H, Jansson S and Shinozaki K (2018) Stable accumulation of photosystem II requires ONE‐HELIX PROTEIN1 (OHP1) of the light harvesting‐like family. Plant Physiol 176, 2277–2291. PubMed PMC

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

Mork‐Jansson A, Bue AK, Gargano D, Furnes C, Reisinger V, Arnold J, Kmiec K and Eichacker LA (2015) Lil3 assembles with proteins regulating chlorophyll synthesis in barley. PLoS ONE 10, e0133145. PubMed PMC

Hey D, Rothbart M, Herbst J, Wang P, Müller J, Wittmann D, Gruhl K and Grimm B (2017) LIL3, a light‐harvesting complex protein, links terpenoid and tetrapyrrole biosynthesis in Arabidopsis thaliana . Plant Physiol 174, 1037–1050. PubMed PMC

Link S, Engelmann K, Meierhoff K and Westhoff P (2012) The atypical short‐chain dehydrogenases HCF173 and HCF244 are jointly involved in translational initiation of the psbA mRNA of Arabidopsis . Plant Physiol 160, 2202–2218. PubMed PMC

Ermakova‐Gerdes S and Vermaas W (1999) Inactivation of the open reading frame slr0399 in Synechocystis sp. PCC 6803 functionally complements mutations near the Q A niche of photosystem II. J Biol Chem 274, 30540–30549. PubMed

Mothersole DJ, Jackson PJ, Vasilev C, Tucker JD, Brindley AA, Dickman MJ and Hunter CN (2016) PucC and LhaA direct efficient assembly of the light‐harvesting complexes in Rhodobacter sphaeroides. Mol Microbiol 99, 307–327. PubMed PMC

Samuelson JC, Chen M, Jiang F, Moller I, Wiedmann M, Kuhn A, Phillips GJ and Dalbey RE (2000) YidC mediates membrane protein insertion in bacteria. Nature 406, 637–641. PubMed

Sobotka R (2014) Making proteins green; biosynthesis of chlorophyll‐binding proteins in cyanobacteria. Photosynth Res 119, 222–232. PubMed

Gerdes L, Bais T, Klostermann E, Karl M, Phillippar K, Hünken M, Soll J and Schünemann D (2006) A second thylakoid membrane‐localized Alb3/OxaI/YidC homologue is involved in proper chloroplast biogenesis in Arabidopsis thaliana . J Biol Chem 281, 16632–16642. PubMed

Bédard J, Trösch R, Wu F, Ling Q, Flores‐Pérez U, Töpel M, Nawarz F and Jarvis P (2017) Suppressors of the chloroplast protein import mutant tic40 reveal a genetic link between protein import and thylakoid biogenesis. Plant Cell 29, 1726–1747. PubMed PMC

Williams JGK (1988) Construction of specific mutations in photosystem II photosynthetic reaction center by genetic engineering methods in Synechocystis 6803. Methods Enzymol 167, 766–778.

Cereda A, Hitchcock A, Symes MD, Cronin L, Bibby TS and Jones AK (2014) A Bioelectrochemical approach to characterize extracellular electron transfer by Synechocystis sp. PCC6803. PLoS ONE 9, e91484. PubMed PMC

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