Structure-function analysis of Lactiplantibacillus plantarum DltE& reveals D-alanylated lipoteichoic acids as direct cues supporting Drosophila juvenile growth
- CNRS UMR 5086, Universite Claude Bernard, France
- Ecole Normale Supérieure de Lyon, France
- Universite Paris-Saclay, INRAE, France
- CNRS UAR3444, INSERM US8, Universite Claude Bernard, France
- Univ Lille, CNRS, UMR 8576, France
Abstract
Metazoans establish mutually beneficial interactions with their resident microorganisms. However, our understanding of the microbial cues contributing to host physiology remains elusive. Previously, we identified a bacterial machinery encoded by the dlt operon involved in Drosophila melanogaster's juvenile growth promotion by Lactiplantibacillus plantarum. Here, using crystallography combined with biochemical and cellular approaches, we investigate the physiological role of an uncharacterized protein (DltE) encoded by this operon. We show that lipoteichoic acids (LTAs) but not wall teichoic acids are D-alanylated in Lactiplantibacillus plantarumNC8 cell envelope and demonstrate that DltE is a D-Ala carboxyesterase removing D-Ala from LTA. Using the mutualistic association of L. plantarumNC8 and Drosophila melanogaster as a symbiosis model, we establish that D-alanylated LTAs (D-Ala-LTAs) are direct cues supporting intestinal peptidase expression and juvenile growth in Drosophila. Our results pave the way to probing the contribution of D-Ala-LTAs to host physiology in other symbiotic models.
Data availability
-Diffraction data have been deposited in PDB under the accession codes 8AGR/8AIK/8AJI/8AKH-All data generated during this study are included in the manuscript and supporting file and a single Source Data file has been provided.
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Crystal structure of DltE from L.plantarum, apo formProtein Data Bank, 8AGR.
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Crystal structure of DltE from L.plantarum, tartare bound formProtein Data Bank, 8AIK.
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Crystal structure of DltE from L.plantarum, TCEP bound formProtein Data Bank, 8AJI.
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Crystal structure of DltE from L.plantarum soaked with LTAProtein Data Bank, 8AKH.
Article and author information
Author details
Funding
Agence Nationale de la Recherche (ANR-18-CE15-0011)
- Nikos Nikolopoulos
- Renata Matos
- Stephanie Ravaud
- Pascal Courtin
- Houssam Akherraz
- Simon Palussiere
- Virginie Gueguen-Chaignon
- Marie Salomon-Mallet
- Alain Guillot
- Yann Guerardel
- Marie-Pierre Chapot-Chartier
- Christophe Grangeasse
- François Leulier
Fondation pour la Recherche Médicale (DEQ20180839196)
- Renata Matos
- Houssam Akherraz
- François Leulier
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2023, Nikolopoulos 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.
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Structure-function analysis of Lactiplantibacillus plantarum DltE& reveals D-alanylated lipoteichoic acids as direct cues supporting Drosophila juvenile growth
eLife 12:e84669.
https://doi.org/10.7554/eLife.84669
Further reading
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- Biochemistry and Chemical Biology
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Teichoic acids (TA) are linear phospho-saccharidic polymers and important constituents of the cell envelope of Gram-positive bacteria, either bound to the peptidoglycan as wall teichoic acids (WTA) or to the membrane as lipoteichoic acids (LTA). The composition of TA varies greatly but the presence of both WTA and LTA is highly conserved, hinting at an underlying fundamental function that is distinct from their specific roles in diverse organisms. We report the observation of a periplasmic space in Streptococcus pneumoniae by cryo-electron microscopy of vitreous sections. The thickness and appearance of this region change upon deletion of genes involved in the attachment of TA, supporting their role in the maintenance of a periplasmic space in Gram-positive bacteria as a possible universal function. Consequences of these mutations were further examined by super-resolved microscopy, following metabolic labeling and fluorophore coupling by click chemistry. This novel labeling method also enabled in-gel analysis of cell fractions. With this approach, we were able to titrate the actual amount of TA per cell and to determine the ratio of WTA to LTA. In addition, we followed the change of TA length during growth phases, and discovered that a mutant devoid of LTA accumulates the membrane-bound polymerized TA precursor.
