Oomycete small RNAs bind to the plant RNA-induced silencing complex for virulence

  1. Florian Dunker
  2. Adriana Trutzenberg
  3. Jan S Rothenpieler
  4. Sarah Kuhn
  5. Reinhard Pröls
  6. Tom Schreiber
  7. Alain Tissier
  8. Ariane Kemen
  9. Eric Kemen
  10. Ralph Hückelhoven
  11. Arne Weiberg  Is a corresponding author
  1. Ludwig Maximilian University of Munich, Germany
  2. Technical University of Munich, Germany
  3. Leibniz Institute of Plant Biochemistry, Germany
  4. University of Tübingen, Germany

Abstract

The exchange of small RNAs (sRNAs) between hosts and pathogens can lead to gene silencing in the recipient organism, a mechanism termed cross-kingdom RNAi (ck-RNAi). While fungal sRNAs promoting virulence are established, the significance of ck-RNAi in distinct plant pathogens is not clear. Here, we describe that sRNAs of the pathogen Hyaloperonospora arabidopsidis, which represents the kingdom of oomycetes and is phylogenetically distant from fungi, employ the host plant's Argonaute (AGO)/RNA-induced silencing complex for virulence. To demonstrate H. arabidopsidis sRNA (HpasRNA) functionality in ck-RNAi, we designed a novel CRISPR endoribonuclease Csy4/GUS reporter that enabled in situ visualization of HpasRNA-induced target suppression in Arabidopsis. The significant role of HpasRNAs together with AtAGO1 in virulence was revealed in plant atago1 mutants and by transgenic Arabidopsis expressing a short-tandem-target-mimic to block HpasRNAs, that both exhibited enhanced resistance. HpasRNA-targeted plant genes contributed to host immunity, as Arabidopsis gene knockout mutants displayed quantitative enhanced susceptibility.

Data availability

Sequencing data have been deposited in NCBI SRA (PRJNA395139).

The following data sets were generated

Article and author information

Author details

  1. Florian Dunker

    Genetics, Ludwig Maximilian University of Munich, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Adriana Trutzenberg

    Genetics, Ludwig Maximilian University of Munich, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Jan S Rothenpieler

    Genetics, Ludwig Maximilian University of Munich, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8892-8230
  4. Sarah Kuhn

    Genetics, Ludwig Maximilian University of Munich, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Reinhard Pröls

    Phytopathology, School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Tom Schreiber

    Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Halle, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Alain Tissier

    Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Halle, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Ariane Kemen

    Center for Plant Molecular Biology, University of Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Eric Kemen

    Center for Plant Molecular Biology, University of Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Ralph Hückelhoven

    Phytopathology, School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany
    Competing interests
    The authors declare that no competing interests exist.
  11. Arne Weiberg

    Genetics, Ludwig Maximilian University of Munich, Martinsried, Germany
    For correspondence
    a.weiberg@lmu.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4300-4864

Funding

Deutsche Forschungsgemeinschaft (WE 5707/1-1)

  • Arne Weiberg

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

Reviewing Editor

  1. Axel A Brakhage, Hans Knöll Institute, Germany

Version history

  1. Received: February 17, 2020
  2. Accepted: May 21, 2020
  3. Accepted Manuscript published: May 22, 2020 (version 1)
  4. Accepted Manuscript updated: May 26, 2020 (version 2)
  5. Version of Record published: June 16, 2020 (version 3)

Copyright

© 2020, Dunker 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

  • 6,072
    views
  • 884
    downloads
  • 87
    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. Florian Dunker
  2. Adriana Trutzenberg
  3. Jan S Rothenpieler
  4. Sarah Kuhn
  5. Reinhard Pröls
  6. Tom Schreiber
  7. Alain Tissier
  8. Ariane Kemen
  9. Eric Kemen
  10. Ralph Hückelhoven
  11. Arne Weiberg
(2020)
Oomycete small RNAs bind to the plant RNA-induced silencing complex for virulence
eLife 9:e56096.
https://doi.org/10.7554/eLife.56096

Share this article

https://doi.org/10.7554/eLife.56096

Further reading

    1. Microbiology and Infectious Disease
    2. Structural Biology and Molecular Biophysics
    Alexander D Cook, Mark Carrington, Matthew K Higgins
    Research Article

    African trypanosomes replicate within infected mammals where they are exposed to the complement system. This system centres around complement C3, which is present in a soluble form in serum but becomes covalently deposited onto the surfaces of pathogens after proteolytic cleavage to C3b. Membrane-associated C3b triggers different complement-mediated effectors which promote pathogen clearance. To counter complement-mediated clearance, African trypanosomes have a cell surface receptor, ISG65, which binds to C3b and which decreases the rate of trypanosome clearance in an infection model. However, the mechanism by which ISG65 reduces C3b function has not been determined. We reveal through cryogenic electron microscopy that ISG65 has two distinct binding sites for C3b, only one of which is available in C3 and C3d. We show that ISG65 does not block the formation of C3b or the function of the C3 convertase which catalyses the surface deposition of C3b. However, we show that ISG65 forms a specific conjugate with C3b, perhaps acting as a decoy. ISG65 also occludes the binding sites for complement receptors 2 and 3, which may disrupt recruitment of immune cells, including B cells, phagocytes, and granulocytes. This suggests that ISG65 protects trypanosomes by combining multiple approaches to dampen the complement cascade.

    1. Microbiology and Infectious Disease
    Michael D Sacco, Lauren R Hammond ... Yu Chen
    Research Article

    In the Firmicutes phylum, GpsB is a membrane associated protein that coordinates peptidoglycan synthesis with cell growth and division. Although GpsB has been studied in several bacteria, the structure, function, and interactome of Staphylococcus aureus GpsB is largely uncharacterized. To address this knowledge gap, we solved the crystal structure of the N-terminal domain of S. aureus GpsB, which adopts an atypical, asymmetric dimer, and demonstrates major conformational flexibility that can be mapped to a hinge region formed by a three-residue insertion exclusive to Staphylococci. When this three-residue insertion is excised, its thermal stability increases, and the mutant no longer produces a previously reported lethal phenotype when overexpressed in Bacillus subtilis. In S. aureus, we show that these hinge mutants are less functional and speculate that the conformational flexibility imparted by the hinge region may serve as a dynamic switch to finetune the function of the GpsB complex and/or to promote interaction with its various partners. Furthermore, we provide the first biochemical, biophysical, and crystallographic evidence that the N-terminal domain of GpsB binds not only PBP4, but also FtsZ, through a conserved recognition motif located on their C-termini, thus coupling peptidoglycan synthesis to cell division. Taken together, the unique structure of S. aureus GpsB and its direct interaction with FtsZ/PBP4 provide deeper insight into the central role of GpsB in S. aureus cell division.