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D-Alanylation of teichoic acids contributes to Lactobacillus plantarum-mediated Drosophila growth during chronic undernutrition

Abstract

The microbial environment influences animal physiology. However, the underlying molecular mechanisms of such functional interactions are largely undefined. Previously, we showed that during chronic undernutrition, strains of Lactobacillus plantarum, a major commensal partner of Drosophila, promote host juvenile growth and maturation partly through enhanced expression of intestinal peptidases. By screening a transposon insertion library of Lactobacillus plantarum in gnotobiotic Drosophila larvae, we identify a bacterial cell-wall-modifying machinery encoded by the pbpX2-dlt operon that is critical to enhance host digestive capabilities and promote animal growth and maturation. Deletion of this operon leads to bacterial cell wall alteration with a complete loss of d-alanylation of teichoic acids. We show that L. plantarum cell walls bearing d-alanylated teichoic acids are directly sensed by Drosophila enterocytes to ensure optimal intestinal peptidase expression and activity, juvenile growth and maturation during chronic undernutrition. We thus conclude that besides peptidoglycan, teichoic acid modifications participate in the host–commensal bacteria molecular dialogue occurring in the intestine.

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Acknowledgements

The authors would like to thank M. Strigini and G. Storelli for critical reading and editing of the manuscript; C. Grangeasse for helpful discussions and help with bacterial cell imaging; C. Login and T. Meylheuc from MIMA2 platform at INRA Jouy-en-Josas Research Center for TEM and SEM sample preparation and observation, respectively; the Arthro-Tools and PLATIM platforms of the SFR Biosciences (UMS3444/US8) for providing Drosophila and imaging facilities; the IGFL sequencing platform for deep sequencing; P. Serror for pG+host9 and H. Licandro-Seraut for the Pjunc-TpaseIS 1223 system. R.C.M. thanks the ‘Fondation pour la Recherche Médicale’ for financial support through a postdoctoral scholarship, SPF20140129318. M.E.M. was funded by the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement N8659510. This work was funded by an ERC starting grant (FP7/2007-2013-No. 309704). The laboratory of F.L. is supported by the FINOVI foundation and the EMBO Young Investigator Program.

Author information

F.L. supervised the work. R.C.M. and F.L. designed the experiments. R.C.M., M.S., A.-L.B., D.M. and H.G. performed the experiments. B.G. and S.H. designed and performed high-throughput insertion tracking by deep sequencing. M.E.M. and P.J. performed the insertion site bioinformatics analysis. P.C. performed d-alanine and PG quantifications. A.-L.B. developed the protocol for proteolytic activity determination. R.C.M., A.-L.B., P.C., M.-P.C.-C., M.S. and F.L. analysed the results. R.C.M. and F.L. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Correspondence to François Leulier.

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Supplementary Table 2

Transposon insertions in coding regions.

Supplementary Table 4

P values for statistical tests.

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Further reading

Fig. 1: Identification of L. plantarum NC8 loci involved in Drosophila growth promotion.
Fig. 2: The pbpX2-dlt operon affects Drosophila’s growth.
Fig. 3: Cell envelope changes related to pbpX2-dlt operon deletion.
Fig. 4: Drosophila-reduced protease expression in the presence of the Δdlt op strain is independent of the Imd pathway.
Fig. 5: Sensing of multiple cell wall motifs is required for Lp NC8-mediated larval growth promotion.