Altered N-glycan composition impacts flagella mediated adhesion in Chlamydomonas reinhardtii

  1. Nannan Xu
  2. Anne Oltmanns
  3. Longsheng Zhao
  4. Antoine Girot
  5. Marzieh Karimi
  6. Lara Hoepfner
  7. Simon Kelterborn
  8. Martin Scholz
  9. Julia Beißel
  10. Peter Hegemann
  11. Oliver Bäumchen
  12. Lu-Ning Liu
  13. Kaiyao Huang  Is a corresponding author
  14. Michael Hippler  Is a corresponding author
  1. Institute of Hydrobiology, Chinese Academy of Sciences, China
  2. University of Münster, Germany
  3. University of Liverpool, United Kingdom
  4. Max Planck Institute for Dynamics and Self-Organization, Germany
  5. Humboldt University of Berlin, Germany
  6. Max Planck Institute for Dynamics and Self Organization, Germany

Abstract

For the unicellular alga Chlamydomonas reinhardtii, the presence of N-glycosylated proteins on the surface of two flagella is crucial for both cell-cell interaction during mating and flagellar surface adhesion. However, it is not known whether only the presence or also the composition of N-glycans attached to respective proteins is important for these processes. To this end, we tested several C. reinhardtii insertional mutants and a CRISPR/Cas9 knockout mutant of xylosyltransferase 1A, all possessing altered N-glycan compositions. Taking advantage of atomic force microscopy and micropipette force measurements, our data revealed that reduction in N-glycan complexity impedes the adhesion force required for binding the flagella to surfaces. This results in impaired polystyrene bead binding and transport but not gliding of cells on solid surfaces. Notably, assembly, intraflagellar transport and protein import into flagella are not affected by altered N-glycosylation. Thus, we conclude that proper N-glycosylation of flagellar proteins is crucial for adhering C. reinhardtii cells onto surfaces, indicating that N-glycans mediate surface adhesion via direct surface contact.

Data availability

The mass spectrometry proteomics data (Figure 1) have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository with the dataset identifier PXD018353.

The following data sets were generated

Article and author information

Author details

  1. Nannan Xu

    Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Anne Oltmanns

    Institute for Plant Biology and Biotechnology, University of Münster, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Longsheng Zhao

    Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  4. Antoine Girot

    Max Planck Institute for Dynamics and Self-Organization, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Marzieh Karimi

    Max Planck Institute for Dynamics and Self-Organization, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Lara Hoepfner

    Institute for Plant Biology and Biotechnology, University of Münster, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8222-4563
  7. Simon Kelterborn

    Institute of Biology, Humboldt University of Berlin, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Martin Scholz

    Biology, University of Münster, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Julia Beißel

    Biology, University of Münster, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Peter Hegemann

    Institute of Biology, Experimental Biophysics, Humboldt University of Berlin, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3589-6452
  11. Oliver Bäumchen

    Max Planck Institute for Dynamics and Self Organization, Goettingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4879-0369
  12. Lu-Ning Liu

    Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8884-4819
  13. Kaiyao Huang

    Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China
    For correspondence
    huangky@ihb.ac.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8669-1065
  14. Michael Hippler

    Institute for Plant Biology and Biotechnology, University of Münster, Münster, Germany
    For correspondence
    mhippler@uni-muenster.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9670-6101

Funding

Deutsche Forschungsgemeinschaft (Hi 737 / 12-1)

  • Michael Hippler

National Natural Science Foundation of China (Grant 31671399)

  • Kaiyao Huang

Royal Society (UF120411,URF\R\180030,RGF\EA\181061 and RGF\EA\180233)

  • Lu-Ning Liu

Biotechnology and Biological Sciences Research Council (BB/R003890/1,BB/M012441/1)

  • Lu-Ning Liu

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

Reviewing Editor

  1. Ahmet Yildiz, University of California, Berkeley, United States

Version history

  1. Received: May 11, 2020
  2. Accepted: December 9, 2020
  3. Accepted Manuscript published: December 10, 2020 (version 1)
  4. Version of Record published: December 24, 2020 (version 2)

Copyright

© 2020, Xu 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

  • 1,740
    views
  • 264
    downloads
  • 10
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

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. Nannan Xu
  2. Anne Oltmanns
  3. Longsheng Zhao
  4. Antoine Girot
  5. Marzieh Karimi
  6. Lara Hoepfner
  7. Simon Kelterborn
  8. Martin Scholz
  9. Julia Beißel
  10. Peter Hegemann
  11. Oliver Bäumchen
  12. Lu-Ning Liu
  13. Kaiyao Huang
  14. Michael Hippler
(2020)
Altered N-glycan composition impacts flagella mediated adhesion in Chlamydomonas reinhardtii
eLife 9:e58805.
https://doi.org/10.7554/eLife.58805

Share this article

https://doi.org/10.7554/eLife.58805

Further reading

    1. Plant Biology
    Ivan Kulich, Julia Schmid ... Jiří Friml
    Research Article

    Root gravitropic bending represents a fundamental aspect of terrestrial plant physiology. Gravity is perceived by sedimentation of starch-rich plastids (statoliths) to the bottom of the central root cap cells. Following gravity perception, intercellular auxin transport is redirected downwards leading to an asymmetric auxin accumulation at the lower root side causing inhibition of cell expansion, ultimately resulting in downwards bending. How gravity-induced statoliths repositioning is translated into asymmetric auxin distribution remains unclear despite PIN auxin efflux carriers and the Negative Gravitropic Response of roots (NGR) proteins polarize along statolith sedimentation, thus providing a plausible mechanism for auxin flow redirection. In this study, using a functional NGR1-GFP construct, we visualized the NGR1 localization on the statolith surface and plasma membrane (PM) domains in close proximity to the statoliths, correlating with their movements. We determined that NGR1 binding to these PM domains is indispensable for NGR1 functionality and relies on cysteine acylation and adjacent polybasic regions as well as on lipid and sterol PM composition. Detailed timing of the early events following graviperception suggested that both NGR1 repolarization and initial auxin asymmetry precede the visible PIN3 polarization. This discrepancy motivated us to unveil a rapid, NGR-dependent translocation of PIN-activating AGCVIII kinase D6PK towards lower PMs of gravity-perceiving cells, thus providing an attractive model for rapid redirection of auxin fluxes following gravistimulation.

    1. Plant Biology
    Daniel S Yu, Megan A Outram ... Simon J Williams
    Research Article

    Plant pathogens secrete proteins, known as effectors, that function in the apoplast or inside plant cells to promote virulence. Effector recognition by cell-surface or cytosolic receptors results in the activation of defence pathways and plant immunity. Despite their importance, our general understanding of fungal effector function and recognition by immunity receptors remains poor. One complication often associated with effectors is their high sequence diversity and lack of identifiable sequence motifs precluding prediction of structure or function. In recent years, several studies have demonstrated that fungal effectors can be grouped into structural classes, despite significant sequence variation and existence across taxonomic groups. Using protein X-ray crystallography, we identify a new structural class of effectors hidden within the secreted in xylem (SIX) effectors from Fusarium oxysporum f. sp. lycopersici (Fol). The recognised effectors Avr1 (SIX4) and Avr3 (SIX1) represent the founding members of the Fol dual-domain (FOLD) effector class, with members containing two distinct domains. Using AlphaFold2, we predicted the full SIX effector repertoire of Fol and show that SIX6 and SIX13 are also FOLD effectors, which we validated experimentally for SIX6. Based on structural prediction and comparisons, we show that FOLD effectors are present within three divisions of fungi and are expanded in pathogens and symbionts. Further structural comparisons demonstrate that Fol secretes effectors that adopt a limited number of structural folds during infection of tomato. This analysis also revealed a structural relationship between transcriptionally co-regulated effector pairs. We make use of the Avr1 structure to understand its recognition by the I receptor, which leads to disease resistance in tomato. This study represents an important advance in our understanding of Fol-tomato, and by extension plant–fungal interactions, which will assist in the development of novel control and engineering strategies to combat plant pathogens.