Cellular characterization of actin gene concerned with contact-dependent mechanisms in Naegleria fowleri.
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.
['Sohn|Hae-Jin|HJ|', 'Song|Kyoung-Ju|KJ|', 'Kang|Heekyoung|H|', 'Ham|A-Jeong|AJ|', 'Lee|Jae-Ho|JH|', 'Chwae|Yong-Joon|YJ|', 'Kim|Kyongmin|K|', 'Park|Sun|S|', 'Kim|Jong-Hyun|JH|', 'Shin|Ho-Joon|HJ|0000-0002-9065-5902']
D000199:Actins D000818:Animals D016466:CHO Cells D020808:Central Nervous System Protozoal Infections D003001:Cloning, Molecular D006224:Cricetinae D003412:Cricetulus D005353:Fibronectins D049452:Green Fluorescent Proteins D006801:Humans D016848:Naegleria fowleri D021381:Protein Transport D053058:Trophozoites
actin gene contact-dependent mechanism Naegleria fowleri phagocytosis adhesion