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Title: A thylakoid membrane-bound and redox-active rubredoxin (RBD1) functions in de novo assembly and repair of photosystem II

Abstract

Photosystem II (PSII) undergoes frequent photooxidative damage that, if not repaired, impairs photosynthetic activity and growth. How photosynthetic organisms protect vulnerable PSII intermediate complexes during de novo assembly and repair remains poorly understood. Here, we report the genetic and biochemical characterization of chloroplast-located rubredoxin 1 (RBD1), a PSII assembly factor containing a redox-active rubredoxin domain and a single C-terminal transmembrane α-helix (TMH) domain. RBD1 is an integral thylakoid membrane protein that is enriched in stroma lamellae fractions with the rubredoxin domain exposed on the stromal side. RBD1 also interacts with PSII intermediate complexes containing cytochromeb559. Complementation of theChlamydomonas reinhardtii(hereafterChlamydomonas) RBD1-deficient2pacmutant with constructs encoding RBD1 protein truncations and site-directed mutations demonstrated that the TMH domain is essential for de novo PSII assembly, whereas the rubredoxin domain is involved in PSII repair. The rubredoxin domain exhibits a redox midpoint potential of +114 mV and is proficient in 1-electron transfers to a surrogate cytochromecin vitro. Reduction of oxidized RBD1 is NADPH dependent and can be mediated by ferredoxin-NADP+reductase (FNR) in vitro. We propose that RBD1 participates, together with the cytochromeb559, in the protection of PSII intermediate complexes from photooxidative damage during de novo assembly and repair. This role of RBD1 is consistent withmore » its evolutionary conservation among photosynthetic organisms and the fact that it is essential in photosynthetic eukaryotes.« less

Authors:
 [1];  [2];  [3];  [4];  [5]; ORCiD logo [6]
  1. Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst., and Dept. of Plant and Microbial Biology
  2. Univ. of California, Berkeley, CA (United States). Dept. of Chemistry
  3. Univ. of California, Berkeley, CA (United States). Dept. of Electron Microscope Laboratory
  4. Ruhr-University Bochum, Bochum (Germany). Molecular Biology of Plant Organelles
  5. Univ. of California, Berkeley, CA (United States). Dept. of Chemistry; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Materials Sciences Division
  6. Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst., and Dept. of Plant and Microbial Biology; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division
Publication Date:
Research Org.:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1561943
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 33; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

García-Cerdán, José G., Furst, Ariel L., McDonald, Kent L., Schünemann, Danja, Francis, Matthew B., and Niyogi, Krishna K. A thylakoid membrane-bound and redox-active rubredoxin (RBD1) functions in de novo assembly and repair of photosystem II. United States: N. p., 2019. Web. doi:10.1073/pnas.1903314116.
García-Cerdán, José G., Furst, Ariel L., McDonald, Kent L., Schünemann, Danja, Francis, Matthew B., & Niyogi, Krishna K. A thylakoid membrane-bound and redox-active rubredoxin (RBD1) functions in de novo assembly and repair of photosystem II. United States. https://doi.org/10.1073/pnas.1903314116
García-Cerdán, José G., Furst, Ariel L., McDonald, Kent L., Schünemann, Danja, Francis, Matthew B., and Niyogi, Krishna K. 2019. "A thylakoid membrane-bound and redox-active rubredoxin (RBD1) functions in de novo assembly and repair of photosystem II". United States. https://doi.org/10.1073/pnas.1903314116. https://www.osti.gov/servlets/purl/1561943.
@article{osti_1561943,
title = {A thylakoid membrane-bound and redox-active rubredoxin (RBD1) functions in de novo assembly and repair of photosystem II},
author = {García-Cerdán, José G. and Furst, Ariel L. and McDonald, Kent L. and Schünemann, Danja and Francis, Matthew B. and Niyogi, Krishna K.},
abstractNote = {Photosystem II (PSII) undergoes frequent photooxidative damage that, if not repaired, impairs photosynthetic activity and growth. How photosynthetic organisms protect vulnerable PSII intermediate complexes during de novo assembly and repair remains poorly understood. Here, we report the genetic and biochemical characterization of chloroplast-located rubredoxin 1 (RBD1), a PSII assembly factor containing a redox-active rubredoxin domain and a single C-terminal transmembrane α-helix (TMH) domain. RBD1 is an integral thylakoid membrane protein that is enriched in stroma lamellae fractions with the rubredoxin domain exposed on the stromal side. RBD1 also interacts with PSII intermediate complexes containing cytochromeb559. Complementation of theChlamydomonas reinhardtii(hereafterChlamydomonas) RBD1-deficient2pacmutant with constructs encoding RBD1 protein truncations and site-directed mutations demonstrated that the TMH domain is essential for de novo PSII assembly, whereas the rubredoxin domain is involved in PSII repair. The rubredoxin domain exhibits a redox midpoint potential of +114 mV and is proficient in 1-electron transfers to a surrogate cytochromecin vitro. Reduction of oxidized RBD1 is NADPH dependent and can be mediated by ferredoxin-NADP+reductase (FNR) in vitro. We propose that RBD1 participates, together with the cytochromeb559, in the protection of PSII intermediate complexes from photooxidative damage during de novo assembly and repair. This role of RBD1 is consistent with its evolutionary conservation among photosynthetic organisms and the fact that it is essential in photosynthetic eukaryotes.},
doi = {10.1073/pnas.1903314116},
url = {https://www.osti.gov/biblio/1561943}, journal = {Proceedings of the National Academy of Sciences of the United States of America},
issn = {0027-8424},
number = 33,
volume = 116,
place = {United States},
year = {Mon Jul 29 00:00:00 EDT 2019},
month = {Mon Jul 29 00:00:00 EDT 2019}
}

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Cited by: 19 works
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Figures / Tables:

Fig. 1 Fig. 1: 2pac is a nonphotosynthetic mutant with impaired assembly of PSII monomers and abnormal chloroplast architecture. (A) Growth phenotype and PSII efficiency (Fv/Fm) of wild-type (WT) and mutant spotted cells grown mixotrophically (TAP) under low light (5 μmol photons·m−2·s−1) or photoautotrophically (HS) under normal light (80 μmol photons·m−2·s−1) conditions.more » (B) Transmission electron microscopy analyses of WT and mutant cells grown heterotrophically. (C) BN-PAGE analyses of WT and 2pac solubilized thylakoid membrane (TM) protein complexes treated with 2 different detergents, $$α$$-DM and $$β$$-DM. (D) The 2D BN/SDS/PAGE and immunoblot analyses from WT and 2pac solubilized TM protein complexes against PSII subunits D2 and CP43, and RBD1. Immunodetection against D2 and CP43 varies. In WT samples, films were recorded after 5-s exposure compared with 1 min in 2pac samples. Lanes are labeled as follows: PSI, photosystem I; PSII-D, PSII dimer; PSII-M, PSII monomer; PSII-SC, PSII–LHCII supercomplexes; uCP43, unassembled CP43. (E) Immunoblots analyses against D2, RBD1, and acetylated tubulin proteins from greening experiment with the chlL mutant, as well as Coomassie-stained SDS/PAGE. Whole-cell samples were collected and subjected to denaturing SDS/PAGE and immunoblot analysis after 0, 6, and 24 h following the shift to normal growth light conditions. One hundred percent loading corresponds to about 1 × 106 cells.« less

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Mechanism of photosystem II photoinactivation and D1 protein degradation at low light: The role of back electron flow
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Works referencing / citing this record:

Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.