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

 Hae-Jin Sohn  ;  Jong-Hyun Kim  ;  Myeong-Heon Shin  ;  Kyoung-Ju Song  ;  Ho-Joon Shin 
 PARASITOLOGY RESEARCH, Vol.106(4) : 917-924, 2010 
Journal Title
Issue Date
Actins/chemistry ; Actins/genetics ; Actins/physiology* ; Amino Acid Sequence ; Animals ; Base Sequence ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Cytoplasm/chemistry ; DNA, Protozoan/chemistry ; DNA, Protozoan/genetics ; Mice ; Mice, Inbred BALB C ; Microscopy, Fluorescence ; Molecular Sequence Data ; Naegleria fowleri/pathogenicity* ; Phagocytosis* ; Protozoan Proteins/chemistry ; Protozoan Proteins/genetics ; Protozoan Proteins/physiology* ; Pseudopodia/chemistry ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Virulence Factors/chemistry ; Virulence Factors/genetics ; Virulence Factors/physiology*
Chinese Hamster Ovary Cell ; Chinese Hamster Ovary ; Antisense Oligonucleotide ; Actin Gene ; Degenerate Oligonucleotide Primer
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.
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1. College of Medicine (의과대학) > Dept. of Environmental Medical Biology (환경의생물학교실) > 1. Journal Papers
Yonsei Authors
Shin, Myeong Heon(신명헌) ORCID logo https://orcid.org/0000-0001-8207-6110
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