title: Phospholipid synthesis in Borrelia burgdorferi: BB0249 and BB0721 encode functional phosphatidylcholine synthase and phosphatidylglycerolphosphate synthase proteins.
abstract: Phospholipids are an important component of bacterial membranes. Borrelia burgdorferi differs from many other bacteria in that it contains only two major membrane phospholipids: phosphatidylglycerol (PG) and phosphatidylcholine (PC). B. burgdorferi appears to lack enzymes required for synthesis of PC through the well-described methylation pathway. However, B. burgdorferi does contain a gene (BB0249) with significant identity to a recently described phosphatidylcholine synthase gene (pcs) of Sinorhizobium meliloti. B. burgdorferi also contains a gene (BB0721) with significant identity to the gene (pgs) encoding phosphatidylglycerolphosphate synthase, an enzyme in the synthetic pathway of PG. Activity of BB0249 was confirmed by cloning the gene into Escherichia coli, which does not produce PC. Transformation with a plasmid carrying BB0249 resulted in production of PC by E. coli, but only in the presence of exogenously supplied choline, as would be predicted for a Pcs. Because loss of Pgs activity is lethal to E. coli, activity of BB0721 was confirmed by the ability of BB0721 to complement an E. coli Pgs(-) mutant. A plasmid containing BB0721 was transformed into a Pgs(-) mutant of E. coli containing a copy of the native gene on a temperature-regulated plasmid. The temperature-regulated plasmid was exchanged for a plasmid containing BB0721 and it was shown that BB0721 was able to replace the lost Pgs function and restore bacterial growth. This study has established the existence and function of two critical enzymes in the synthesis of PC and PG in B. burgdorferi. Understanding of the biosynthetic pathways of PC and PG in B. burgdorferi is the first step in delineating the role of these phospholipids in the pathogenesis of Lyme disease.
authors.affiliation: Tupper Research Institute, Division of Geographic Medicine and Infectious Diseases, New England Medical Center, Tufts University School of Medicine, Box 41, 750 Washington Street, Boston, MA 02111, USA.
authors.forename: Xing-Guo
authors.initials: XG
authors.lastname: Wang
authors.affiliation: Tupper Research Institute, Division of Geographic Medicine and Infectious Diseases, New England Medical Center, Tufts University School of Medicine, Box 41, 750 Washington Street, Boston, MA 02111, USA.
authors.forename: Joanna P
authors.initials: JP
authors.lastname: Scagliotti
authors.affiliation: Tupper Research Institute, Division of Geographic Medicine and Infectious Diseases, New England Medical Center, Tufts University School of Medicine, Box 41, 750 Washington Street, Boston, MA 02111, USA.
authors.forename: Linden T
authors.initials: LT
authors.lastname: Hu
mesh_terms: D001483:Base Sequence; D025065:Borrelia burgdorferi / Q000201:enzymology* / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D007700:Kinetics; D010743:Phospholipids / Q000096:biosynthesis*; D010957:Plasmids / Q000235:genetics; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics* / Q000378:metabolism
chemical_list: D010743:Phospholipids; D017855:Transferases (Other Substituted Phosphate Groups); C483298:CDP-diacylglycerol choline O-phosphatidyltransferase; C020676:CDP-diacylglycerol-glycerol-3-phosphate 3-phosphatidyltransferase