title: A set of glycosylphosphatidyl inositol-anchored membrane proteins of Plasmodium falciparum is refractory to genetic deletion.
abstract: Targeted gene disruption has proved to be a powerful approach for studying the function of important ligands involved in erythrocyte invasion by the extracellular merozoite form of the human malaria parasite, Plasmodium falciparum. Merozoite invasion proceeds via a number of seemingly independent alternate pathways, such that entry can proceed with parasites lacking particular ligand-receptor interactions. To date, most focus in this regard has been on single-pass (type 1) membrane proteins that reside in the secretory organelles. Another class of merozoite proteins likely to include ligands for erythrocyte receptors are the glycosylphosphatidyl inositol (GPI)-anchored membrane proteins that coat the parasite surface and/or reside in the apical organelles. Several of these are prominent vaccine candidates, although their functions remain unknown. Here, we systematically attempted to disrupt the genes encoding seven of the known GPI-anchored merozoite proteins of P. falciparum by using a double-crossover gene-targeting approach. Surprisingly, and in apparent contrast to other merozoite antigen classes, most of the genes (six of seven) encoding GPI-anchored merozoite proteins are refractory to genetic deletion, with the exception being the gene encoding merozoite surface protein 5 (MSP-5). No distinguishable growth rate or invasion pathway phenotype was detected for the msp-5 knockout line, although its presence as a surface-localized protein was confirmed.
authors.affiliation: Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC 3050, Australia.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Sanders
authors.forename: Lev M
authors.initials: LM
authors.lastname: Kats
authors.forename: Damien R
authors.initials: DR
authors.lastname: Drew
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: O'Donnell
authors.forename: Matthew
authors.initials: M
authors.lastname: O'Neill
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Maier
authors.forename: Ross L
authors.initials: RL
authors.lastname: Coppel
authors.forename: Brendan S
authors.initials: BS
authors.lastname: Crabb
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D017353:Gene Deletion; D017261:Glycosylphosphatidylinositols / Q000737:chemistry* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010963:Plasmodium falciparum / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D000953:Antigens, Protozoan; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D015800:Protozoan Proteins; C118839:merozoite surface protein 4, Plasmodium