The role of the Psb28 protein in the structure and function of the photosystem II (PSII) complex has been studied in the cyanobacterium Synechocystis sp. PCC 6803. The protein was localized in the membrane fraction and, whereas most of the protein was detected as an unassembled protein, a small portion was found in the PSII core complex lacking the CP43 antenna (RC47). The association of Psb28 with RC47 was further confirmed by preferential isolation of RC47 from the strain containing a histidine-tagged derivative of Psb28 using nickel-affinity chromatography. However, the affinity-purified fraction also contained a small amount of the unassembled PSII inner antenna CP47 bound to Psb28-histidine, indicating a structural relationship between Psb28 and CP47. A psb28 deletion mutant exhibited slower autotrophic growth than wild type, although the absence of Psb28 did not affect the functional properties of PSII. The mutant showed accelerated turnover of the D1 protein, faster PSII repair, and a decrease in the cellular content of PSI. Radioactive labeling revealed a limitation in the synthesis of both CP47 and the PSI subunits PsaA/PsaB in the absence of Psb28. The mutant cells contained a high level of magnesium protoporphyrin IX methylester, a decreased level of protochlorophyllide, and released large quantities of protoporphyrin IX into the medium, indicating inhibition of chlorophyll (Chl) biosynthesis at the cyclization step yielding the isocyclic ring E. Overall, our results show the importance of Psb28 for synthesis of Chls and/or apoproteins of Chl-binding proteins CP47 and PsaA/PsaB.

Institute of Microbiology Academy of Sciences 37981 Trebon Czech Republic

Zobrazit více v PubMed

Ajlani G, Vernotte C (1998) Construction and characterization of a phycobiliprotein-less mutant of Synechocystis sp. PCC 6803. Plant Mol Biol 37 577–580 PubMed

Barber J (2006) Photosystem II: an enzyme of global significance. Biochem Soc Trans 34 619–631 PubMed

Bumba L, Tichý M, Dobáková M, Komenda J, Vácha F (2005) Localization of the PsbH subunit in photosystem II from the Synechocystis 6803 using the His-tagged Ni-NTA nanogold labeling. J Struct Biol 152 28–35 PubMed

Burke E, Barik S (2003) Megaprimer PCR: application in mutagenesis and gene fusion. Methods Mol Biol 226 525–532 PubMed

Dobáková M, Tichý M, Komenda J (2007) Role of the PsbI protein in Photosystem II assembly and repair in the cyanobacterium Synechocystis sp. PCC 6803. Plant Physiol 145 1681–1691 PubMed PMC

Ferreira KN, Iverson TN, Maglaoui K, Barber J, Iwata S (2004) Architecture of the photosynthetic oxygen-evolving center. Science 303 1831–1838 PubMed

Irrgang KD, Shi LX, Funk Ch, Schröder WP (1995) A nuclear-encoded subunit of the photosystem II reaction center. J Biol Chem 270 17588–17593 PubMed

Kashino Y, Lauber WM, Carroll JA, Wang Q, Whitmarsh J, Satoh K, Pakrasi HB (2002) Proteomic analysis of a highly active photosystem II preparation from the cyanobacterium Synechocystis sp. PCC 6803 reveals the presence of novel polypeptides. Biochemistry 41 8004–8012 PubMed

Komenda J, Barber J (1995) Comparison of psbO and psbH deletion mutants of Synechocystis PCC 6803 indicates that degradation of D1 protein is regulated by the QB site and is dependent on protein synthesis. Biochemistry 34 9625–9631 PubMed

Komenda J, Barker M, Kuviková S, DeVries R, Mullineaux CW, Tichý M, Nixon PJ (2006) The FtsH protease slr0228 is important for quality control of Photosystem II in the thylakoid membrane of Synechocystis PCC 6803. J Biol Chem 281 1145–1151 PubMed

Komenda J, Lupínková L, Kopecký J (2002) Absence of the psbH gene product destabilizes the photosystem II complex and bicarbonate binding on its acceptor side in Synechocystis PCC 6803. Eur J Biochem 269 610–619 PubMed

Komenda J, Reisinger V, Müller BC, Dobáková M, Granvogl B, Eichacker LA (2004) Accumulation of the D2 protein is a key regulatory step for assembly of the photosystem II reaction center complex in Synechocystis PCC 6803. J Biol Chem 279 48620–48629 PubMed

Komenda J, Tichý M, Eichacker L (2005) The PsbH protein is associated with the inner antenna CP47 and facilitates D1 processing and incorporation into Photosystem II in the cyanobacterium Synechocystis PCC 6803. Plant Cell Physiol 46 1477–1483 PubMed

Kuviková S, Tichý M, Komenda J (2005) A role of the C-terminal extension of the Photosystem II D1 protein in sensitivity of the cyanobacterium Synechocystis PCC 6803 to photoinhibition. Photochem Photobiol Sci 4 1044–1048 PubMed

Lagarde D, Beuf L, Vermaas W (2000) Increased production of zeaxanthin and other pigments by application of genetic engineering techniques to Synechocystis sp. strain PCC 6803. Appl Environ Microbiol 66 64–72 PubMed PMC

Loll B, Kern J, Saenger W, Zouni A, Biesiadka J (2005) Towards complete cofactor arrangement in the 3.0 Å resolution structure of photosystem II. Nature 438 1040–1044 PubMed

Lorković ZJ, Schröder WP, Pakrasi HB, Irrgang KD, Herrmann RG, Oelmüller R (1995) Molecular characterization of PsbW, a nuclear-encoded component of the photosystem II reaction center complex in spinach. Proc Natl Acad Sci USA 92 8930–8934 PubMed PMC

Meskauskiene R, Nater M, Goslings D, Kessler F, Op den Camp R, Apel K (2001) FLU: A negative regulator of chlorophyll biosynthesis in Arabidopsis thaliana. Proc Natl Acad Sci USA 98 12826–12831 PubMed PMC

Minamizaki K, Mizoguchi T, Goto T, Tamiaki H, Fujita Y (2008) Identification of two homologous genes, chlAI and chlAII, that are differentially involved in isocyclic ring formation of chlorophyll a in the cyanobacterium Synechocystis sp. PCC 6803. J Biol Chem 283 2684–2692 PubMed

Nanba O, Satoh K (1987) Isolation of a photosystem II reaction center consisting of D-1 and D-2 polypeptides and cytochrome b-559. Proc Natl Acad Sci USA 84 109–112 PubMed PMC

Pakrasi HB, Williams JGK, Arntzen CJ (1988) Targeted mutagenesis of the psbE and psbF blocks photosynthetic electron transport: evidence for a functional role of cytochrome b-559 in photosystem II. EMBO J 7 325–332 PubMed PMC

Promnares K, Komenda J, Bumba L, Nebesarova J, Vacha F, Tichy 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 PubMed

Rokka A, Suorsa M, Saleem A, Battchikova N, Aro EM (2005) Synthesis and assembly of thylakoid protein complex: multiple assembly steps of photosystem II. Biochem J 388 159–168 PubMed PMC

Sakurai I, Mizusawa N, Ohashi S, Kobayashi M, Wada H (2007) Effects of the lack of phosphatidylglycerol on the donor side of photosystem II. Plant Physiol 144 1336–1346 PubMed PMC

Schägger H, von Jagow G (1991) Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal Biochem 199 223–231 PubMed

Shi LX, Lorković ZJ, Oelmüller R, Schröder WP (2000) The low molecular mass PsbW protein is involved in the stabilization of the dimeric photosystem II complex in Arabidopsis thaliana. J Biol Chem 275 37945–37950 PubMed

Shi LX, Schröder WP (2004) The low molecular mass subunits of the photosynthetic supracomplex, photosystem II. Biochim Biophys Acta 1608 75–96 PubMed

Sobotka R, Dühring U, Komenda J, Peter E, Gardian Z, Tichy M, Grimm B, Wilde A (2008) Importance of the cyanobacterial Gun4 protein for chlorophyll metabolism and assembly of photosynthetic complexes. J Biol Chem 283 25794–25802 PubMed PMC

Tichý M, Lupínková L, Sicora C, Vass I, Kuviková S, Prášil O, Komenda J (2003) Synechocystis 6803 mutants expressing distinct forms of the photosystem II D1 protein from Synechococcus 7942: Relationship between the psbA coding region and sensitivity to visible and UV-B radiation. Biochim Biophys Acta 1605 55–66 PubMed

Tottey S, Block MA, Allen M, Westergren T, Albrieux C, Schiller HV, Merchant S, Jensen PE (2003) Arabidopsis CHL27, located in both envelope and thylakoid membranes, is required for the synthesis of protochlorophyllide. Proc Natl Acad Sci USA 100 16119–16124 PubMed PMC

Vavilin D, Yao D, Vermaas W (2007) Small cab-like proteins retard degradation of Photosystem II-associated chlorophyll in Synechocystis sp. PCC 6803. J Biol Chem 282 306–314 PubMed

Vermaas WFJ, Ikeuchi M, Inoue Y (1988) Protein composition of the photosystem II core complex in genetically engineered mutants of the cyanobacterium Synechocystis sp. PCC 6803. Photosynth Res 17 97–113 PubMed

Wellburn AR, Lichtenthaler K (1984) Formulae and programme to determine total carotenoids and chlorophyll a and b of leaf extracts in different solvents. In C Sybesma, ed, Advances in Photosynthesis Research. Martinus Nijjhoff, Dordrecht, The Netherlands, pp 10–12

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

Yao D, Kieselbach T, Komenda J, Promnares K, Hernandez 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 PubMed

Enhancing the production of chlorophyll f in the cyanobacterium Synechocystis sp. PCC 6803

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

Assembly of D1/D2 complexes of photosystem II: Binding of pigments and a network of auxiliary proteins

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

High-light-inducible proteins HliA and HliB: pigment binding and protein-protein interactions

2.4-Å structure of the double-ring Gemmatimonas phototrophica photosystem

Mutations Suppressing the Lack of Prepilin Peptidase Provide Insights Into the Maturation of the Major Pilin Protein in Cyanobacteria

The Photosystem II Assembly Factor Ycf48 from the Cyanobacterium Synechocystis sp. PCC 6803 Is Lipidated Using an Atypical Lipobox Sequence

Evolution of Ycf54-independent chlorophyll biosynthesis in cyanobacteria

Fast Diffusion of the Unassembled PetC1-GFP Protein in the Cyanobacterial Thylakoid Membrane

Purification of Protein-complexes from the Cyanobacterium Synechocystis sp. PCC 6803 Using FLAG-affinity Chromatography

Chlorophyll f synthesis by a super-rogue photosystem II complex

A Photosynthesis-Specific Rubredoxin-Like Protein Is Required for Efficient Association of the D1 and D2 Proteins during the Initial Steps of Photosystem II Assembly

Sequential deletions of photosystem II assembly factors Ycf48, Ycf39 and Pam68 result in progressive loss of autotrophy in the cyanobacterium Synechocystis PCC 6803

New Urea Derivatives Are Effective Anti-senescence Compounds Acting Most Likely via a Cytokinin-Independent Mechanism

Ycf48 involved in the biogenesis of the oxygen-evolving photosystem II complex is a seven-bladed beta-propeller protein

The cyanobacterial protoporphyrinogen oxidase HemJ is a new b-type heme protein functionally coupled with coproporphyrinogen III oxidase

Binding of pigments to the cyanobacterial high-light-inducible protein HliC

The Ribosome-Bound Protein Pam68 Promotes Insertion of Chlorophyll into the CP47 Subunit of Photosystem II