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Phosphatidylcholine (PC) is the most abundant type of phospholipids in eukaryotes constituting ~30% of total lipids in  Leishmania . PC synthesis mainly occurs  via  the choline branch of the Kennedy pathway (choline ⇒ choline-phosphate ⇒ CDP-choline ⇒ PC) and the N-methylation of phosphatidylethanolamine (PE). In addition,  Leishmania  parasites can acquire PC and other lipids from the host or culture medium. In this study, we assessed the function and essentiality of choline ethanolamine phosphotransferase (CEPT) in  Leishmania major  which is responsible for the final step of the  de novo  synthesis of PC and PE. Our data indicate that CEPT is localized in the endoplasmic reticulum and possesses the activity to generate PC from CDP-choline and diacylglycerol. Targeted deletion of  CEPT  is only possible in the presence of an episomal  CEPT  gene in the promastigote stage of  L. major . These chromosomal null parasites require the episomal expression of  CEPT  to survive in culture, confirming its essentiality during the promastigote stage. In contrast, during  in vivo  infection of BALB/c mice, these chromosomal null parasites appeared to lose the episomal copy of  CEPT  while maintaining normal levels of virulence, replication and cellular PC. Therefore, while the  de novo  synthesis of PC/PE is indispensable for the proliferation of promastigotes, intracellular amastigotes appear to acquire most of their lipids through salvage and remodeling.

De Novo Synthesis of Phosphatidylcholine Is Essential for the Promastigote But Not Amastigote Stage in Leishmania major