1. Microbiology and Infectious Disease
  2. Structural Biology and Molecular Biophysics
Download icon

Malaria parasites use a soluble RhopH complex for erythrocyte invasion and an integral form for nutrient uptake

  1. Marc A Schureck
  2. Joseph E Darling
  3. Alan Merk
  4. Jinfeng Shao
  5. Geervani Daggupati
  6. Prakash Srinivasan
  7. Paul D B Olinares
  8. Michael P Rout
  9. Brian T Chait
  10. Kurt Wollenberg
  11. Sriram Subramaniam  Is a corresponding author
  12. Sanjay A Desai  Is a corresponding author
  1. National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States
  2. National Cancer Institute, NIH, United States
  3. Johns Hopkins Bloomberg School of Public Health, United States
  4. The Rockefeller University, United States
  5. University of British Columbia, Canada
Research Article
  • Cited 1
  • Views 580
  • Annotations
Cite this article as: eLife 2021;10:e65282 doi: 10.7554/eLife.65282

Abstract

Malaria parasites use the RhopH complex for erythrocyte invasion and channel-mediated nutrient uptake. As the member proteins are unique to Plasmodium spp., how they interact and traffic through subcellular sites to serve these essential functions is unknown. We show that RhopH is synthesized as a soluble complex of CLAG3, RhopH2, and RhopH3 with 1:1:1 stoichiometry. After transfer to a new host cell, the complex crosses a vacuolar membrane surrounding the intracellular parasite and becomes integral to the erythrocyte membrane through a PTEX translocon-dependent process. We present a 2.9 Å single-particle cryo-electron microscopy structure of the trafficking complex, revealing that CLAG3 interacts with the other subunits over large surface areas. This soluble complex is tightly assembled with extensive disulfide bonding and predicted transmembrane helices shielded. We propose a large protein complex stabilized for trafficking but poised for host membrane insertion through large-scale rearrangements, paralleling smaller two-state pore-forming proteins in other organisms.

Article and author information

Author details

  1. Marc A Schureck

    Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, United States
    Competing interests
    No competing interests declared.
  2. Joseph E Darling

    Laboratory of Cell Biology, National Cancer Institute, NIH, Bethesda, United States
    Competing interests
    No competing interests declared.
  3. Alan Merk

    Laboratory of Cell Biology, National Cancer Institute, NIH, Bethesda, United States
    Competing interests
    No competing interests declared.
  4. Jinfeng Shao

    Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, United States
    Competing interests
    No competing interests declared.
  5. Geervani Daggupati

    Department of Molecular Microbiology and Immunology, and Johns Hopkins Malaria Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, United States
    Competing interests
    No competing interests declared.
  6. Prakash Srinivasan

    Department of Molecular Microbiology and Immunology, and Johns Hopkins Malaria Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, United States
    Competing interests
    No competing interests declared.
  7. Paul D B Olinares

    Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3429-6618
  8. Michael P Rout

    Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  9. Brian T Chait

    Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  10. Kurt Wollenberg

    Office of Cyber Infrastructure & Computational Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, United States
    Competing interests
    No competing interests declared.
  11. Sriram Subramaniam

    Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada
    For correspondence
    Sriram.Subramaniam@ubc.ca
    Competing interests
    Sriram Subramaniam, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4231-4115
  12. Sanjay A Desai

    Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, United States
    For correspondence
    sdesai@niaid.nih.gov
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2150-2483

Funding

National Institute of Allergy and Infectious Diseases

  • Sanjay A Desai

National Cancer Institute

  • Sriram Subramaniam

National Institutes of Health (P41 GM103314)

  • Brian T Chait

National Institutes of Health (P41 GM109824)

  • Michael P Rout
  • Brian T Chait

Canada Excellence Research Chairs, Government of Canada

  • Sriram Subramaniam

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

Reviewing Editor

  1. Olivier Silvie, Sorbonne Université, UPMC Univ Paris 06, INSERM, CNRS, France

Publication history

  1. Received: November 29, 2020
  2. Accepted: January 4, 2021
  3. Accepted Manuscript published: January 4, 2021 (version 1)
  4. Accepted Manuscript updated: January 5, 2021 (version 2)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 580
    Page views
  • 198
    Downloads
  • 1
    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)

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

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

Further reading

    1. Ecology
    2. Microbiology and Infectious Disease
    Lara Urban et al.
    Research Article

    While traditional microbiological freshwater tests focus on the detection of specific bacterial indicator species, including pathogens, direct tracing of all aquatic DNA through metagenomics poses a profound alternative. Yet, in situ metagenomic water surveys face substantial challenges in cost and logistics. Here, we present a simple, fast, cost-effective and remotely accessible freshwater diagnostics workflow centred around the portable nanopore sequencing technology. Using defined compositions and spatiotemporal microbiota from surface water of an example river in Cambridge (UK), we provide optimised experimental and bioinformatics guidelines, including a benchmark with twelve taxonomic classification tools for nanopore sequences. We find that nanopore metagenomics can depict the hydrological core microbiome and fine temporal gradients in line with complementary physicochemical measurements. In a public health context, these data feature relevant sewage signals and pathogen maps at species level resolution. We anticipate that this framework will gather momentum for new environmental monitoring initiatives using portable devices.

    1. Microbiology and Infectious Disease
    Philip P Adams et al.
    Research Article

    Many bacterial genes are regulated by RNA elements in their 5´ untranslated regions (UTRs). However, the full complement of these elements is not known even in the model bacterium Escherichia coli. Using complementary RNA-sequencing approaches, we detected large numbers of 3´ ends in 5´ UTRs and open reading frames (ORFs), suggesting extensive regulation by premature transcription termination. We documented regulation for multiple transcripts, including spermidine induction involving Rho and translation of an upstream ORF for an mRNA encoding a spermidine efflux pump. In addition to discovering novel sites of regulation, we detected short, stable RNA fragments derived from 5´ UTRs and sequences internal to ORFs. Characterization of three of these transcripts, including an RNA internal to an essential cell division gene, revealed that they have independent functions as sRNA sponges. Thus, these data uncover an abundance of cis- and trans-acting RNA regulators in bacterial 5´ UTRs and internal to ORFs.