Localization of the phosphoethanolamine methyltransferase of the human malaria parasite Plasmodium falciparum to the Golgi apparatus.
Phosphatidylcholine is the most abundant phospholipid in the membranes of Plasmodium falciparum, the agent of severe human malaria. The synthesis of this phospholipid occurs via two routes, the CDP-choline pathway, which uses host choline as a precursor, and the plant-like serine decarboxylase-phosphoethanolamine methyltransferase (SDPM) pathway, which uses host serine as a precursor. Although various components of these pathways have been identified, their cellular locations remain unknown. We have previously reported the identification and characterization of the phosphoethanolamine methyltransferase, Pfpmt, of P. falciparum and shown that it plays a critical role in the synthesis of phosphatidylcholine via the SDPM pathway. Here we provide the first evidence that the transmethylation step of the SDPM pathway occurs in the parasite Golgi apparatus. We show that the level of Pfpmt protein in the infected erythrocyte is regulated in a stage-specific fashion, with high levels detected during the trophozoite stage at the peak of parasite membrane biogenesis. Confocal microscopy revealed that Pfpmt is not cytoplasmic. Immunoelectron microscopy revealed that Pfpmt localizes to membrane structures that extend from the nuclear membrane but that it only partially co-localizes with the endoplasmic reticulum marker BiP. Using transgenic parasites expressing green fluorescent protein targeted to different cellular compartments, a complete co-localization was detected with Rab6, a marker of the Golgi apparatus. Together these studies provide the first evidence that the transmethylation step of the SDPM pathway of P. falciparum occurs in the Golgi apparatus and indicate an important role for this organelle in parasite membrane biogenesis.
Witola
William H
WH
Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut 06030-3301.
Pessi
Gabriella
G
Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut 06030-3301.
El Bissati
Kamal
K
Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut 06030-3301.
Reynolds
Jennifer M
JM
Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut 06030-3301.
Mamoun
Choukri Ben
CB
Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut 06030-3301. Electronic address: Choukri@up.uchc.edu.
D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry; D003566:Cytidine Diphosphate Choline / Q000737:chemistry; D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology*; D006056:Golgi Apparatus / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D008780:Methyltransferases / Q000096:biosynthesis*; D010963:Plasmodium falciparum / Q000201:enzymology*; D015800:Protozoan Proteins / Q000737:chemistry; D012694:Serine / Q000737:chemistry; D019076:Transgenes; D020691:rab GTP-Binding Proteins / Q000378:metabolism
D000953:Antigens, Protozoan; D015800:Protozoan Proteins; C094135:Rab6 protein; D012694:Serine; D003566:Cytidine Diphosphate Choline; D008780:Methyltransferases; C102844:phosphoethanolamine methyltransferase; D020691:rab GTP-Binding Proteins