TY - JOUR
T1 - Characterization of inositol lipid metabolism in gut-associated Bacteroidetes
AU - Heaver, Stacey L.
AU - Le, Henry H.
AU - Tang, Peijun
AU - Baslé, Arnaud
AU - Mirretta Barone, Claudia
AU - Vu, Dai Long
AU - Waters, Jillian L.
AU - Marles-Wright, Jon
AU - Johnson, Elizabeth L.
AU - Campopiano, Dominic J.
AU - Ley, Ruth E.
N1 - Funding Information:
This work was supported by the Max Planck Society. We thank K. Hipp and J. Berger of the Electron Microscopy Core Facility at the Max Planck Institute for Biology Tübingen for their expert imaging of the bacterial capsules; A. Goodman (Yale School of Medicine, USA) for providing relevant strains of B. thetaiotaomicron; D. Warschawski for training on fatty acid methyl ester analysis; the Diamond Light Source (Oxfordshire, UK) for beamtime (proposal mx24948) and staff of beamline I04. D.J.C. and J.M.-W. acknowledge the funding provided by the Biotechnology and Biological Sciences Research Council (BBSRC, grants BB/V001620/1 and BB/V00168X/1). This work was supported in part by NIH grant R24GM137782-01 to Parastoo Azadi of the Complex Carbohydrate Research Center at the University of Georgia.
Funding Information:
This work was supported by the Max Planck Society. We thank K. Hipp and J. Berger of the Electron Microscopy Core Facility at the Max Planck Institute for Biology Tübingen for their expert imaging of the bacterial capsules; A. Goodman (Yale School of Medicine, USA) for providing relevant strains of B. thetaiotaomicron; D. Warschawski for training on fatty acid methyl ester analysis; the Diamond Light Source (Oxfordshire, UK) for beamtime (proposal mx24948) and staff of beamline I04. D.J.C. and J.M.-W. acknowledge the funding provided by the Biotechnology and Biological Sciences Research Council (BBSRC, grants BB/V001620/1 and BB/V00168X/1). This work was supported in part by NIH grant R24GM137782-01 to Parastoo Azadi of the Complex Carbohydrate Research Center at the University of Georgia.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/6/20
Y1 - 2022/6/20
N2 - Inositol lipids are ubiquitous in eukaryotes and have finely tuned roles in cellular signalling and membrane homoeostasis. In Bacteria, however, inositol lipid production is relatively rare. Recently, the prominent human gut bacterium Bacteroides thetaiotaomicron (BT) was reported to produce inositol lipids and sphingolipids, but the pathways remain ambiguous and their prevalence unclear. Here, using genomic and biochemical approaches, we investigated the gene cluster for inositol lipid synthesis in BT using a previously undescribed strain with inducible control of sphingolipid synthesis. We characterized the biosynthetic pathway from myo-inositol-phosphate (MIP) synthesis to phosphoinositol dihydroceramide, determined the crystal structure of the recombinant BT MIP synthase enzyme and identified the phosphatase responsible for the conversion of bacterially-derived phosphatidylinositol phosphate (PIP-DAG) to phosphatidylinositol (PI-DAG). In vitro, loss of inositol lipid production altered BT capsule expression and antimicrobial peptide resistance. In vivo, loss of inositol lipids decreased bacterial fitness in a gnotobiotic mouse model. We identified a second putative, previously undescribed pathway for bacterial PI-DAG synthesis without a PIP-DAG intermediate, common in Prevotella. Our results indicate that inositol sphingolipid production is widespread in host-associated Bacteroidetes and has implications for symbiosis.
AB - Inositol lipids are ubiquitous in eukaryotes and have finely tuned roles in cellular signalling and membrane homoeostasis. In Bacteria, however, inositol lipid production is relatively rare. Recently, the prominent human gut bacterium Bacteroides thetaiotaomicron (BT) was reported to produce inositol lipids and sphingolipids, but the pathways remain ambiguous and their prevalence unclear. Here, using genomic and biochemical approaches, we investigated the gene cluster for inositol lipid synthesis in BT using a previously undescribed strain with inducible control of sphingolipid synthesis. We characterized the biosynthetic pathway from myo-inositol-phosphate (MIP) synthesis to phosphoinositol dihydroceramide, determined the crystal structure of the recombinant BT MIP synthase enzyme and identified the phosphatase responsible for the conversion of bacterially-derived phosphatidylinositol phosphate (PIP-DAG) to phosphatidylinositol (PI-DAG). In vitro, loss of inositol lipid production altered BT capsule expression and antimicrobial peptide resistance. In vivo, loss of inositol lipids decreased bacterial fitness in a gnotobiotic mouse model. We identified a second putative, previously undescribed pathway for bacterial PI-DAG synthesis without a PIP-DAG intermediate, common in Prevotella. Our results indicate that inositol sphingolipid production is widespread in host-associated Bacteroidetes and has implications for symbiosis.
U2 - 10.1038/s41564-022-01152-6
DO - 10.1038/s41564-022-01152-6
M3 - Article
C2 - 35725777
AN - SCOPUS:85132159679
SN - 2058-5276
VL - 7
SP - 986
EP - 1000
JO - Nature Microbiology
JF - Nature Microbiology
IS - 7
ER -