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The Nf-actin gene is an important factor for food-cup formation and cytotoxicity of pathogenic Naegleria fowleri.

Sohn, HJ; Kim, JH; Shin, MH; Song, KJ; Shin, HJ
Parasitology research, 106(4):917-924, 2010
Journal Title
Parasitology research
Naegleria fowleri destroys target cells by trogocytosis, a phagocytosis mechanism, and a process of piecemeal ingestion of target cells by food-cups. Phagocytosis is an actin-dependent process that involves polymerization of monomeric G-actin into filamentous F-actin. However, despite the numerous studies concerning phagocytosis, its role in the N. fowleri food-cup formation related with trogocytosis has been poorly reported. In this study, we cloned and characterized an Nf-actin gene to elucidate the role of Nf-actin gene in N. fowleri pathogenesis. The Nf-actin gene is composed of 1,128-bp and produced a 54.1-kDa recombinant protein (Nf-actin). The sequence identity was 82% with nonpathogenic Naegleria gruberi but has no sequence identity with other mammals or human actin gene. Anti-Nf-actin polyclonal antibody was produced in BALB/c mice immunized with recombinant Nf-actin. The Nf-actin was localized on the cytoplasm, pseudopodia, and especially, food-cup structure (amoebastome) in N. fowleri trophozoites using immunofluorescence assay. When N. fowleri co-cultured with Chinese hamster ovary cells, Nf-actin was observed to localize around on phagocytic food-cups. We also observed that N. fowleri treated with cytochalasin D as actin polymerization inhibitor or transfected with antisense oligomer of Nf-actin gene had shown the reduced ability of food-cup formation and in vitro cytotoxicity. Finally, it suggests that Nf-actin plays an important role in phagocytic activity of pathogenic N. fowleri.
MeSH terms
Actins/chemistryActins/geneticsActins/physiology*Amino Acid SequenceAnimalsBase SequenceCHO CellsCloning, MolecularCricetinaeCricetulusCytoplasm/chemistryDNA, Protozoan/chemistryDNA, Protozoan/geneticsMiceMice, Inbred BALB CMicroscopy, FluorescenceMolecular Sequence DataNaegleria fowleri/pathogenicity*Phagocytosis*Protozoan Proteins/chemistryProtozoan Proteins/geneticsProtozoan Proteins/physiology*Pseudopodia/chemistrySequence Analysis, DNASequence Homology, Amino AcidVirulence Factors/chemistryVirulence Factors/geneticsVirulence Factors/physiology*
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
AJOU Authors
김, 종현신, 호준
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