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Parasite Immunol. 2019 May 11:e12631. doi: 10.1111/pim.12631. [Epub ahead of print]

Cellular characterization of actin gene concerned with contact-dependent mechanisms in Naegleria fowleri.

Sohn HJ1,2, Song KJ3, Kang H1,2, Ham AJ1,2, Lee JH1,2, Chwae YJ1,2, Kim K1,2, Park S1,2, Kim JH4, Shin HJ1,2.

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Department of Microbiology, Ajou University School of Medicine, Suwon, Korea.
Department of Biomedical Science, Graduate School of Ajou University, Suwon, Korea.
PurgoFarm Co. Ltd, Hwasung, Gyeonggi, Korea.
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju, Korea.


Free-living amoeba, Naegleria fowleri, destroys target cells through contact-dependent mechanisms, such as phagocytosis and/or trogocytosis. A previous experiment showed that the nf-actin gene consisted of 1.2 kbp, produced a 50.1 kDa recombinant protein (Nf-actin), and was localized on the cytoskeleton, pseudopodia and amoebastome. In this study, cellular characterization of the nf-actin gene concerned with contact-dependent mechanisms in N fowleri was performed. The nf-actin gene was amplified from a gene-cloned vector, pEXQP5-T7/NT TOPO. The nf-actin gene was introduced into the Ubi-pEGFP-C2 vector, and Ubi-pEGFP-C2/nf-actin was transfected into N fowleri trophozoites. Strong GFP fluorescence was detected in N fowleri trophozoites transfected with Ubi-pEGFP-C2/nf-actin. Expression of EGFP-Nf-actin protein was detected by Western blot analysis. The nf-actin-overexpressing N fowleri showed significantly increased adhesion activity against extracellular matrix components, fibronectin, collagen I and fibrinogen, compared with wild-type N fowleri. Moreover, nf-actin-overexpressing N fowleri showed increased phagocytic activity and cytotoxicity in comparison with wild-type N fowleri. In summary, the overexpressed nf-actin gene has an important function in ability to increase cell adhesion, cytotoxicity and phagocytosis by N fowleri.


Naegleria fowleri ; actin gene; adhesion; contact-dependent mechanism; phagocytosis


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