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Characterization of a Synechocystis sp. PCC 6803 double mutant lacking the CyanoP and Ycf48 proteins of Photosystem II

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Abstract

Homologs of the Photosystem II (PS II) subunit PsbP are found in plants, algae, and cyanobacteria. In higher plants, PsbP is associated with mature PS II centers, but in cyanobacteria, the homologous CyanoP protein appears sub-stoichiometric to PS II. We have investigated the role of CyanoP by characterizing knockout mutants of the cyanobacterium Synechocystis sp. PCC 6803. Removal of CyanoP resulted in changes to phycobilisome coupling and energy transfer to PS II, but the function of PS II itself remained similar to wild type. We therefore investigated the hypothesis that CyanoP is involved in the biogenesis or repair of PS II by creating a double mutant lacking both CyanoP and the PS II assembly factor Ycf48. This strain exhibited an additive reduction in the amplitude of variable chlorophyll a fluorescence induction relative to either of the single mutants but displayed increased oxygen evolution, slight increases in PS II monomer and dimer levels, and a reduction in accumulation of an early PS II assembly complex containing CP47, compared to the ΔYcf48 strain.

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Abbreviations

Bis–Tris:

2-[Bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)propane-1,3-diol

BMF:

Blue measuring flashes

BN-PAGE:

Blue-native polyacrylamide gel electrophoresis

CP43:

43-kDa chlorophyll a-binding protein of the core antenna

CP47:

47-kDa chlorophyll a-binding protein of the core antenna

D1:

Photosystem II reaction center protein subunit

D2:

Photosystem II reaction center protein subunit

DCMU:

3-(3,4-Dichlorophenyl)-1,1-dimethylurea

FO :

Dark-adapted (minimum) fluorescence

FM :

Maximum level of chlorophyll a fluorescence

J and I:

Inflection points between O (FO) and P in the chlorophyll a fluorescence induction curve

LC/MS/MS:

Liquid chromatography tandem mass spectrometry

LHC:

Light-harvesting complex

O:

Origin (FO) level of the chlorophyll a fluorescence induction curve

OD:

Optical density

P:

Peak level in the chlorophyll a fluorescence induction curve

PS I:

Photosystem I

PS II:

Photosystem II

QA :

Primary plastoquinone electron acceptor of PS II

QB :

Secondary plastoquinone electron acceptor of PS II

RC47:

Reaction center complex lacking the CP43 protein

RMF:

Red measuring flashes

S-states:

Oxidation states of the oxygen-evolving complex of PS II

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

Tris:

2-Amino-2-hydroxymethyl-propane-1,3-diol

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Acknowledgments

S.A.J. was supported by a University of Otago Division of Health Sciences Career Development Postdoctoral Fellowship. Other funding for this project was provided by an Otago University Research grant to J.E.-R. We thank Asher J. Dale for assistance with preliminary experiments.

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  1. Department of Biochemistry, University of Otago, Dunedin, 9016, New Zealand

    Simon A. Jackson & Julian J. Eaton-Rye

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  1. Simon A. Jackson
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Correspondence to Julian J. Eaton-Rye.

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Jackson, S.A., Eaton-Rye, J.J. Characterization of a Synechocystis sp. PCC 6803 double mutant lacking the CyanoP and Ycf48 proteins of Photosystem II. Photosynth Res 124, 217–229 (2015). https://doi.org/10.1007/s11120-015-0122-0

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  • DOI: https://doi.org/10.1007/s11120-015-0122-0

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