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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.

Author information

1
Department of Microbiology, Ajou University School of Medicine, Suwon, Korea.
2
Department of Biomedical Science, Graduate School of Ajou University, Suwon, Korea.
3
PurgoFarm Co. Ltd, Hwasung, Gyeonggi, Korea.
4
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju, Korea.

Abstract

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.

KEYWORDS:

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

PMID:
31077592
DOI:
10.1111/pim.12631

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