Analysis of the psbU Gene Encoding the 12-kDa Extrinsic Protein of Photosystem II and Studies on Its Role by Deletion Mutagenesis in Synechocystis sp. PCC 6803

Shen, Jian Ren; Ikeuchi, Masahiko; Inoue, Yorinao

doi:10.1074/jbc.272.28.17821

Cited by 77 publications

(87 citation statements)

References 30 publications

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“…The Control and the Site-directed Mutated Cyt c 550 -Shen and Inoue (5) suggested that all the cyt c 550 present in the cells was attached to the PS II. Here, however, we presented data suggesting that in control whole cells, in addition to the PS II-bound cyt c 550 , a significant fraction of cyt c 550 is present in the soluble form.…”

Section: Discussionmentioning

confidence: 99%

Cytochrome c550 in the Cyanobacterium Thermosynechococcus elongatus

Kirilovsky

Roncel

Boussac

et al. 2004

Journal of Biological Chemistry

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Cytochrome c 550 is one of the extrinsic Photosystem II subunits in cyanobacteria and red algae. To study the possible role of the heme of the cytochrome c 550 we constructed two mutants of Thermosynechococcus elongatus in which the residue His-92, the sixth ligand of the heme, was replaced by a Met or a Cys in order to modify the redox properties of the heme. The H92M and H92C mutations changed the midpoint redox potential of the heme in the isolated cytochrome by ؉125 mV and ؊30 mV, respectively, compared with the wild type. The binding-induced increase of the redox potential observed in the wild type and the H92C mutant was absent in the H92M mutant. Both modified cytochromes were more easily detachable from the Photosystem II compared with the wild type. The Photosystem II activity in cells was not modified by the mutations suggesting that the redox potential of the cytochrome c 550 is not important for Photosystem II activity under normal growth conditions. A mutant lacking the cytochrome c 550 was also constructed. It showed a lowered affinity for Cl ؊ and Ca 2؉ as reported earlier for the cytochrome c 550 -less Synechocystis 6803 mutant, but it showed a shorter lived S 2 Q B Ϫ state, rather than a stabilized S 2 state and rapid deactivation of the enzyme in the dark, which were characteristic of the Synechocystis mutant. It is suggested that the latter effects may be caused by loss (or weaker binding) of the other extrinsic proteins rather than a direct effect of the absence of the cytochrome c 550 .Cytochrome c 550 (cyt c 550 ), 1 present in cyanobacteria and red algae, was first discovered by Holton and Myers (1) as a soluble monoheme c-type cytochrome. The cyt c 550 has a molecular mass of about 15 kDa, His/His coordination and a very low redox potential around Ϫ260 mV (for review see Ref.2). showed that cyt c 550 is stoichiometrically bound to the Photosystem II (PS II), activates oxygen evolving activity and allows the binding of the 12 kDa protein, another extrinsic component of the cyanobacterial PS II involved in oxygen evolution. The three-dimensional structure of the PS II from two thermophilic cyanobacteria strains confirmed that cyt c 550 binds to the luminal PS II surface in the vicinity of the D1 and CP43 proteins (6 -8). The phenotype of the ⌬psbV (cyt c 550 -less) mutant of Synechocystis PCC 6803 was already characterized with respect to PS II activity (9 -11). The ⌬psbV and the double ⌬psbV/⌬psbU (encoding the 12 kDa protein) mutants were unable to grow in the absence of Ca 2ϩ and Cl Ϫ ions, their PS II activity decreased to 40% of the wild type, and they showed a very rapid inactivation of the enzyme in the dark (after 2 h, the activity decreased about 90% whereas in the wild type only 25% of the activity was lost). A slight retardation in O 2 release from the S 3 state was also observed in these mutants. Another effect observed in these Synechocystis PCC 6803 mutants was a large intensity decrease of the B band of thermoluminescence (TL) and an upshift in the temperature maxima of the ...

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Section: Discussionmentioning

confidence: 99%

Cytochrome c550 in the Cyanobacterium Thermosynechococcus elongatus

Kirilovsky

Roncel

Boussac

et al. 2004

Journal of Biological Chemistry

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“…Synechocystis 6803 cells were prepared as described earlier (27). SDS-PAGE was performed as described in Ref.…”

Section: Sds-page and Western Blot Analysis Of Mans Protein-membranesmentioning

confidence: 99%

A Two-component Signal Transduction Pathway Regulates Manganese Homeostasis in Synechocystis 6803, a Photosynthetic Organism

Ogawa

Bao

Katoh

et al. 2002

Journal of Biological Chemistry

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Elemental manganese is essential for the production of molecular oxygen by cyanobacteria, plants, and algae. In the cyanobacterium Synechocystis sp. PCC 6803, transcription of the mntCAB operon, encoding a high affinity Mn transporter, occurs under Mn starvation (nM Mn) conditions but not in Mn-sufficient (M Mn) growth medium. Using a strain in which the promoter of this operon directs the transcription of the luxAB reporter genes, we determined that inactivation of the slr0640 gene, which encodes a histidine kinase sensor protein component of a two-component signal transduction system, resulted in constitutive high levels of lux luminescence. Systematic targeted inactivation mutagenesis also identified slr1837 as the gene encoding the corresponding response regulator protein. We have named these two genes manS (manganese-sensor) and manR (manganese-regulator), respectively. A polyhistidinetagged form of the ManS protein was localized in the Synechocystis 6803 cell membrane. Directed replacement of the conserved catalytic His-205 residue of this protein by Leu abolished its activity, although the mutated protein was present in cyanobacterial membrane. This mutant also showed suboptimal rates of Mn uptake under either Mn-starved or Mn-sufficient growth condition. These data suggest that the ManS/ManR two-component system plays a central role in the homeostasis of manganese in Synechocystis 6803 cells.

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“…Middle phase Slow phase Non-decaying phase From our flash fluorescence data we can conclude that forward electron transport from It has been demonstrated in previous studies that inactivation of the psbU gene resulted mostly in structural destabilization of the donor side of PSII (Shen et al, 1997;Shen et al, 1998;Nishiyama et al, 1999). However, our data show, in agreement with previous findings (Veerman 612 2 et al, 2005), that the absence of the PsbU subunit modifies not only the OEC, but also the acceptor side components of PSII.…”

Section: Fast Phasementioning

confidence: 85%

“…PsbV can also bind independently, but it is stabilized by presence of PsbO and PsbU. In contrast, PsbU cannot bind in the absence of PsbO or PsbV (Shen and Inoue, 1993;Shen et al, 1997;Eaton-Rye et al, 2003). Thus it is thought that PsbO binds first, then PsbV, and PsbU is the last to bind.…”

Section: Presence Of the Ocp Gene In Diferent Cyanobacteriamentioning

confidence: 99%

“…The absence of PsbU results 352 2 in the reduction of the rate of oxygen evolution (Shen et al, 1997;Inoue-Kashino et al, 2005) and a high sensitivity of cells to photodamage, resulting in rapid degradation of the D1 protein (Inoue-Kashino et al, 2005;Balint et al, 2006 (Shen et al, 1997). Shen et al (1997) demonstrated that the peak temperatures of both the thermoluminescence Q and B bands were increased by 4 o C in ∆PsbU Synechocystis mutant cells compared to wild type cells. The presence of PsbU contributes to the heat stability of the oxygen-evolving machinery during the acclimation to high temperature (Nishiyama et al, 1997;Nishiyama et al, 1999).…”

Section: The Effect Of Psbu At the Donor And Acceptor Side Of Psiimentioning

confidence: 99%

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The role of the orange carotenoid protein and PsbU protein in photoprotection in cyanobacteria

Abasova¹

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Analysis of the psbU Gene Encoding the 12-kDa Extrinsic Protein of Photosystem II and Studies on Its Role by Deletion Mutagenesis in Synechocystis sp. PCC 6803

Cited by 77 publications

References 30 publications

Cytochrome c550 in the Cyanobacterium Thermosynechococcus elongatus

Cytochrome c550 in the Cyanobacterium Thermosynechococcus elongatus

A Two-component Signal Transduction Pathway Regulates Manganese Homeostasis in Synechocystis 6803, a Photosynthetic Organism

The role of the orange carotenoid protein and PsbU protein in photoprotection in cyanobacteria

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