Identification of distinct pH-and zeaxanthin-dependent quenching in LHCSR3 from Chlamydomonas reinhardtii

  1. Julianne M Troiano
  2. Federico Perozeni
  3. Raymundo Moya
  4. Luca Zuliani
  5. Kwangyrul Baek
  6. EonSeon Jin
  7. Stefano Cazzaniga
  8. Matteo Ballottari  Is a corresponding author
  9. Gabriela S Schlau-Cohen  Is a corresponding author
  1. Massachusetts Institute of Technology, United States
  2. University of Verona, Italy
  3. Hanyang University, Republic of Korea

Abstract

Under high light, oxygenic photosynthetic organisms avoid photodamage by thermally dissipating absorbed energy, which is called non-photochemical quenching. In green algae, a chlorophyll and carotenoid-binding protein, light-harvesting complex stress-related (LHCSR3), detects excess energy via a pH drop and serves as a quenching site. Using a combined in vivo and in vitro approach, we investigated quenching within LHCSR3 from Chlamydomonas reinhardtii. In vitro two distinct quenching processes, individually controlled by pH and zeaxanthin, were identified within LHCSR3. The pH-dependent quenching was removed within a mutant LHCSR3 that lacks the residues that are protonated to sense the pH drop. Observation of quenching in zeaxanthin-enriched LHCSR3 even at neutral pH demonstrated zeaxanthin-dependent quenching, which also occurs in other light-harvesting complexes. Either pH- or zeaxanthin-dependent quenching prevented the formation of damaging reactive oxygen species, and thus the two quenching processes may together provide different induction and recovery kinetics for photoprotection in a changing environment.

Data availability

Source data files have been provided for Figures 1A and 3.Single-molecule photon emission data for Figures 1B-D and 2 has been deposited on Zenodo.org and is available at 10.5281/zenodo.4306869

The following data sets were generated

Article and author information

Author details

  1. Julianne M Troiano

    Department of Chemistry, Massachusetts Institute of Technology, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Federico Perozeni

    Department of Biotechnology, University of Verona, Verona, Italy
    Competing interests
    The authors declare that no competing interests exist.
  3. Raymundo Moya

    Department of Chemistry, Massachusetts Institute of Technology, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Luca Zuliani

    Department of Biotechnology, University of Verona, Verona, Italy
    Competing interests
    The authors declare that no competing interests exist.
  5. Kwangyrul Baek

    Department of Life Science, Hanyang University, Seoul, Republic of Korea
    Competing interests
    The authors declare that no competing interests exist.
  6. EonSeon Jin

    Department of Life Science, Hanyang University, Seoul, Republic of Korea
    Competing interests
    The authors declare that no competing interests exist.
  7. Stefano Cazzaniga

    Department of Biotechnology, University of Verona, Verona, Italy
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2824-7916
  8. Matteo Ballottari

    Department of Biotechnology, University of Verona, Verona, Italy
    For correspondence
    matteo.ballottari@univr.it
    Competing interests
    The authors declare that no competing interests exist.
  9. Gabriela S Schlau-Cohen

    Department of Chemistry, Massachusetts Institute of Technology, Cambridge, United States
    For correspondence
    gssc@mit.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7746-2981

Funding

Human Frontiers Science Program (RGY0076)

  • Gabriela S Schlau-Cohen

National Science Foundation (CHE-1740645)

  • Gabriela S Schlau-Cohen

H2020 European Research Council (679814)

  • Matteo Ballottari

Korea Ministry of Science and ICT (NRF-2014M1A8A1049273)

  • EonSeon Jin

Arnold and Mabel Beckman Foundation (Postdoctoral Fellowship)

  • Julianne M Troiano

National Science Foundation (Graduate Research Fellowship)

  • Raymundo Moya

Arnold and Mabel Beckman Foundation (Beckman Young Investigator)

  • Gabriela S Schlau-Cohen

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. David M Kramer, Michigan State University, United States

Publication history

  1. Received: June 24, 2020
  2. Accepted: January 14, 2021
  3. Accepted Manuscript published: January 15, 2021 (version 1)
  4. Version of Record published: February 5, 2021 (version 2)

Copyright

© 2021, Troiano et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,742
    Page views
  • 264
    Downloads
  • 13
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Julianne M Troiano
  2. Federico Perozeni
  3. Raymundo Moya
  4. Luca Zuliani
  5. Kwangyrul Baek
  6. EonSeon Jin
  7. Stefano Cazzaniga
  8. Matteo Ballottari
  9. Gabriela S Schlau-Cohen
(2021)
Identification of distinct pH-and zeaxanthin-dependent quenching in LHCSR3 from Chlamydomonas reinhardtii
eLife 10:e60383.
https://doi.org/10.7554/eLife.60383

Further reading

    1. Plant Biology
    Josephine HR Maidment, Motoki Shimizu ... Mark J Banfield
    Research Article

    A subset of plant intracellular NLR immune receptors detect effector proteins, secreted by phytopathogens to promote infection, through unconventional integrated domains which resemble the effector’s host targets. Direct binding of effectors to these integrated domains activates plant defenses. The rice NLR receptor Pik-1 binds the Magnaporthe oryzae effector AVR-Pik through an integrated heavy metal-associated (HMA) domain. However, the stealthy alleles AVR-PikC and AVR-PikF avoid interaction with Pik-HMA and evade host defenses. Here, we exploited knowledge of the biochemical interactions between AVR-Pik and its host target, OsHIPP19, to engineer novel Pik-1 variants that respond to AVR-PikC/F. First, we exchanged the HMA domain of Pikp-1 for OsHIPP19-HMA, demonstrating that effector targets can be incorporated into NLR receptors to provide novel recognition profiles. Second, we used the structure of OsHIPP19-HMA to guide the mutagenesis of Pikp-HMA to expand its recognition profile. We demonstrate that the extended recognition profiles of engineered Pikp-1 variants correlate with effector binding in planta and in vitro, and with the gain of new contacts across the effector/HMA interface. Crucially, transgenic rice producing the engineered Pikp-1 variants was resistant to blast fungus isolates carrying AVR-PikC or AVR-PikF. These results demonstrate that effector target-guided engineering of NLR receptors can provide new-to-nature disease resistance in crops.

    1. Cell Biology
    2. Plant Biology
    Han Nim Lee, Jenu Varghese Chacko ... Marisa S Otegui
    Research Article Updated

    The ubiquitin-binding NBR1 autophagy receptor plays a prominent role in recognizing ubiquitylated protein aggregates for vacuolar degradation by macroautophagy. Here, we show that upon exposing Arabidopsis plants to intense light, NBR1 associates with photodamaged chloroplasts independently of ATG7, a core component of the canonical autophagy machinery. NBR1 coats both the surface and interior of chloroplasts, which is then followed by direct engulfment of the organelles into the central vacuole via a microautophagy-type process. The relocalization of NBR1 into chloroplasts does not require the chloroplast translocon complexes embedded in the envelope but is instead greatly enhanced by removing the self-oligomerization mPB1 domain of NBR1. The delivery of NBR1-decorated chloroplasts into vacuoles depends on the ubiquitin-binding UBA2 domain of NBR1 but is independent of the ubiquitin E3 ligases SP1 and PUB4, known to direct the ubiquitylation of chloroplast surface proteins. Compared to wild-type plants, nbr1 mutants have altered levels of a subset of chloroplast proteins and display abnormal chloroplast density and sizes upon high light exposure. We postulate that, as photodamaged chloroplasts lose envelope integrity, cytosolic ligases reach the chloroplast interior to ubiquitylate thylakoid and stroma proteins which are then recognized by NBR1 for autophagic clearance. This study uncovers a new function of NBR1 in the degradation of damaged chloroplasts by microautophagy.