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

• Published in: Parasite immunology Vol. 41; no. 8; pp. e12631 - n/a
• Main Authors: Sohn, Hae‐Jin, Song, Kyoung‐Ju, Kang, Heekyoung, Ham, A‐Jeong, Lee, Jae‐Ho, Chwae, Yong‐Joon, Kim, Kyongmin, Park, Sun, Kim, Jong‐Hyun, Shin, Ho‐Joon
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.08.2019
• Subjects: Actin; actin gene; Actins - genetics; Actins - metabolism; adhesion; Amoeba; Animals; Cell adhesion; Cell adhesion & migration; Central Nervous System Protozoal Infections - genetics; Central Nervous System Protozoal Infections - metabolism; Central Nervous System Protozoal Infections - parasitology; CHO Cells; Cloning, Molecular; Collagen (type I); contact‐dependent mechanism; Cricetinae; Cricetulus; Cytoskeleton; Cytotoxicity; Extracellular matrix; Fibrinogen; Fibronectin; Fibronectins - genetics; Fibronectins - metabolism; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Humans; Naegleria fowleri; Naegleria fowleri - genetics; Naegleria fowleri - growth & development; Naegleria fowleri - metabolism; Phagocytes; Phagocytosis; Plasma membranes; Protein Transport; Pseudopodia; Trophozoites; Trophozoites - genetics; Trophozoites - growth & development; Trophozoites - metabolism; phagocytosis; Naegleria fowleri; adhesion; contact-dependent mechanism; actin gene
• ISSN: 0141-9838; 1365-3024
• DOI: 10.1111/pim.12631

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.