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Characterization of photosystem II assembly complexes containing ONE-HELIX PROTEIN1 in Arabidopsis thaliana

Abstract

The assembly process of photosystem II (PSII) requires several auxiliary proteins to form assembly intermediates. In plants, early assembly intermediates comprise D1 and D2 subunits of PSII together with a few auxiliary proteins including at least ONE-HELIX PROTEIN1 (OHP1), OHP2, and HIGH-CHLOROPHYLL FLUORESCENCE 244 (HCF244) proteins. Herein, we report the basic characterization of the assembling intermediates, which we purified from Arabidopsis transgenic plants overexpressing a tagged OHP1 protein and named the OHP1 complexes. We analyzed two major forms of OHP1 complexes by mass spectrometry, which revealed that the complexes consist of OHP1, OHP2, and HCF244 in addition to the PSII subunits D1, D2, and cytochrome b559. Analysis of chlorophyll fluorescence showed that a major form of the complex binds chlorophyll a and carotenoids and performs quenching with a time constant of 420 ps. To identify the localization of the auxiliary proteins, we solubilized thylakoid membranes using a digitonin derivative, glycodiosgenin, and separated them into three fractions by ultracentrifugation, and detected these proteins in the loose pellet containing the stroma lamellae and the grana margins together with two chlorophyll biosynthesis enzymes. The results indicated that chlorophyll biosynthesis and assembly may take place in the same compartments of thylakoid membranes. Inducible suppression of the OHP2 mRNA substantially decreased the OHP2 protein in mature Arabidopsis leaves without a significant reduction in the maximum quantum yield of PSII under low-light conditions, but it compromised the yields under high-light conditions. This implies that the auxiliary protein is required for acclimation to high-light conditions.

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Acknowledgements

The authors would like to thank Ms. Junko Kishimoto for her excellent technical assistance. We also thank Dr. Seiko Oka from the Open Facility, Global Facility Center, Creative Research Institution, Hokkaido University for allowing us to conduct mass spectroscopic analysis of protein samples using LTQ Orbitrap Discovery and providing insight and expertise that greatly assisted with our research. We thank Dr. Yusuke Kato (Okayama University) for providing the anti-D1 antibody, and Dr. Yao-Pin Lin and Dr. Yee-yung Charng (Academia Sinica, Taiwan) for providing the anti-ChlG antibody.

Funding

This study was supported by funding from the Japan Society for the Promotion of Science (JSPS) through the KAKENHI Grant numbers 16H06554 and 20H03017 to RT, 21K06217 to SIO, and 16H06553 and 20H03017 to SA, respectively.

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Contributions

Conceptualization: RT; Experimentation: HM, KT, YU, KS, AY, KY, S-IO, HI, SA, AT; Writing—original draft preparation: HM and RT; Writing—review and editing: HM, SA, RT; Funding acquisition: S-IO, SA, RT; Resources: FM, KS; Supervision: YT, AT, RT.

Corresponding author

Correspondence to Ryouichi Tanaka.

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The authors declare that they have no conflict of interest.

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Maeda, H., Takahashi, K., Ueno, Y. et al. Characterization of photosystem II assembly complexes containing ONE-HELIX PROTEIN1 in Arabidopsis thaliana. J Plant Res 135, 361–376 (2022). https://doi.org/10.1007/s10265-022-01376-x

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  • DOI: https://doi.org/10.1007/s10265-022-01376-x

Keywords

  • Arabidopsis
  • Chlorophyll
  • One-helix proteins
  • Photosystem II