TY - JOUR
T1 - De novo synthesis of phosphatidylcholine is essential for the promastigote but not amastigote stage in Leishmania major
AU - Moitra, Samrat
AU - Basu, Somrita
AU - Pawlowic, Mattie
AU - Hsu, Fong-Fu
AU - Zhang, Kai
N1 - Funding Information:
We thank Dr. Jay Bangs (University at Buffalo, SUNY) for providing the rabbit anti-T. brucei BiP antiserum and Veronica Hernandez (Texas Tech University) for technical assistance.
Funding Information:
This work was supported by the US National Institutes of Health grants AI099380 (KZ), P41-GM103422 (FH), P60-DK20579 (FH), and P30-DK56341 (FH) for the Biomedical Mass Spectrometry Resource at Washington University in St. Louis, MO, USA). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Publisher Copyright:
© Copyright © 2021 Moitra, Basu, Pawlowic, Hsu and Zhang.
PY - 2021/3/12
Y1 - 2021/3/12
N2 - 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.
AB - 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.
KW - amastigote
KW - host
KW - phospholipid
KW - protozoan
KW - salvage
UR - http://www.scopus.com/inward/record.url?scp=85103341595&partnerID=8YFLogxK
U2 - 10.3389/fcimb.2021.647870
DO - 10.3389/fcimb.2021.647870
M3 - Article
C2 - 33777852
VL - 11
JO - Frontiers in Cellular and Infection Microbiology
JF - Frontiers in Cellular and Infection Microbiology
M1 - 647870
ER -