Phosphatidic acid as a limiting host metabolite for the proliferation of the microsporidium Tubulinosema ratisbonensis in Drosophila flies


Microsporidia are located at the base of the fungal evolutionary tree. They are obligate intracellular parasites and harness host metabolism to fuel their growth and proliferation. However, how the infestation of cells affects the whole organism and how the organism contributes to parasite proliferation remain poorly understood. Here, we have developed a Tubulinosema ratisbonensis systemic infection model in the genetically amenable Drosophila melanogaster host, in which parasite spores obtained in a mammalian cell culture infection system are injected into adult flies. The parasites proliferate within flies and ultimately kill their hosts. As commonly observed for microsporidia infecting insects, T.ratisbonensis preferentially grows in the fat body and ultimately depletes the host metabolic stores. We find that supplementing the fly diet with yeast does not benefit the host but the parasite, which increases its proliferation. Unexpectedly, fatty acids and not carbohydrates or amino acids are the critical components responsible for this phenomenon. Our genetic dissection of host lipid metabolism identifies a crucial compound hijacked by T.ratisbonensis: phosphatidic acid. We propose that phosphatidic acid is a limiting precursor for the synthesis of the parasite membranes and, hence, of its proliferation.

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Fig. 1: T.ratisbonensis infection alters D.melanogaster fitness.
Fig. 2: T.ratisbonensis proliferates in the fat body at the expense of host lipids.
Fig. 3: Fatty acid as the limiting factor for the proliferation of T.ratisbonensis.
Fig. 4: Fatty acid availability in the diet determines the proliferation of T.ratisbonensis.
Fig. 5: Phosphatidic acid is a compound scavenged by T.ratisbonensis that limits its proliferation in Drosophila.
Fig. 6: Synthetic view of the pathways beneficial or detrimental to the proliferation of T.ratisbonensis in D.melanogaster.

Data availability

The data sets, including the 16S rRNA DNA sequences of bacteria retrieved in the haemolymph of control or infected flies, generated and/or analysed during the current study are available in the ‘Phosphatidic acid as a limiting host metabolite for the proliferation of the microsporidium T.ratisbonensis in Drosophila flies’ repository:


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We are grateful to M. Yamba and M.-C. Lacombe for expert technical assistance. We thank F. Delbac, A. Gould, B. Lemaitre and P. Léopold for informative discussions and suggestions, and S. Liégeois for critical reading of the manuscript. Transgenic fly stocks were obtained from the VDRC ( Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. This work has been funded by CNRS, University of Strasbourg, Fondation pour la Recherche Médicale (Equipe FRM to D.F. and FDT20140930952 to A.F.), Agence Nationale de la Recherche (BEELOSS, ANR-11-EQPX-0022) and the Institut d’Etude Avancée de l’Université de Strasbourg.

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A.F. performed most of the experimental work described in this study, with assistance from G.C. for the natural infestation experiment and the ingestion/injection of phosphatidic acid into control or microsporidia-infected flies. S.N. developed the infection model and all techniques required to maintain parasite cultures, as well as those for infection of flies; he participated in all aspects of the early stages of this study. A.F. and D.F. designed the experiments, analysed the data and wrote the manuscript.

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Correspondence to Dominique Ferrandon.

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Franchet, A., Niehus, S., Caravello, G. et al. Phosphatidic acid as a limiting host metabolite for the proliferation of the microsporidium Tubulinosema ratisbonensis in Drosophila flies. Nat Microbiol 4, 645–655 (2019).

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