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<p><i>Naegleria fowleri</i> is a free-living amoeba found in lakes, soil, hot springs, and poorly chlorinated swimming pools. It is pathogenic to humans, causing a rare and fatal brain infection known as primary amoebic meningoencephalitis (PAM). A previous study utilized RNA-seq analysis to examine genes expressed in <i>N. fowleri</i> cysts and trophozoites, focusing on the <i>nf-profilin</i> gene, which showed high expression in cysts. Profilin is a small actin-binding protein that regulates <i>nf-actin</i> polymerization and cell movement. Sequence analysis revealed 83% similarity with non-pathogenic <i>N. gruberi</i> and 38% similarity with <i>Acanthamoeba castellanii</i>. Nf-profilin was found to be associated with <i>N. fowleri</i> lysates but not with lysates from other amoebae, as shown by Western blot analysis. Immunofluorescence assays demonstrated that <i>nf-profilin</i> primarily localized to the cell membrane in <i>N. fowleri</i> cysts, while <i>nf-actin</i> localized to the cytoplasm, pseudopodia, and food-cup structures. Real-time RT-PCR indicated higher expression of the <i>nf-profilin</i> gene in cysts compared to trophozoites. In co-culture experiments with target cells, Nf-profilin was initially expressed in the cytoplasm of <i>N. fowleri</i> cysts and the morphology of cyst gradually transitioned to the trophozoite form. Concurrently, the expression of Nf-profilin protein decreased, while Nf-actin protein began to appear in the pseudopodia and food-cups of trophozoites. In conclusion, the <i>nf-profilin</i> and <i>nf-actin</i> genes exhibited complementary expression patterns based on the life stage of <i>N. fowleri</i>, indicating their critical roles in the survival and proliferation. This study emphasizes the significance of actin-binding proteins in understanding the infection and pathogenic mechanisms of <i>N. fowleri</i>.</p>

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Cloning of nf-profilin and intercellular interaction with nf-actin in Naegleria fowleri cysts

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年2月27日

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  • Hae-Jin Sohn,
  • A-Jeong Ham,
  • A-Young Park,
  • Jeong-Heon Lee,
  • Sun Park,
  • Ho-Joon Shin,
  • Jong-Hyun Kim

摘要

Naegleria fowleri is a free-living amoeba found in lakes, soil, hot springs, and poorly chlorinated swimming pools. It is pathogenic to humans, causing a rare and fatal brain infection known as primary amoebic meningoencephalitis (PAM). A previous study utilized RNA-seq analysis to examine genes expressed in N. fowleri cysts and trophozoites, focusing on the nf-profilin gene, which showed high expression in cysts. Profilin is a small actin-binding protein that regulates nf-actin polymerization and cell movement. Sequence analysis revealed 83% similarity with non-pathogenic N. gruberi and 38% similarity with Acanthamoeba castellanii. Nf-profilin was found to be associated with N. fowleri lysates but not with lysates from other amoebae, as shown by Western blot analysis. Immunofluorescence assays demonstrated that nf-profilin primarily localized to the cell membrane in N. fowleri cysts, while nf-actin localized to the cytoplasm, pseudopodia, and food-cup structures. Real-time RT-PCR indicated higher expression of the nf-profilin gene in cysts compared to trophozoites. In co-culture experiments with target cells, Nf-profilin was initially expressed in the cytoplasm of N. fowleri cysts and the morphology of cyst gradually transitioned to the trophozoite form. Concurrently, the expression of Nf-profilin protein decreased, while Nf-actin protein began to appear in the pseudopodia and food-cups of trophozoites. In conclusion, the nf-profilin and nf-actin genes exhibited complementary expression patterns based on the life stage of N. fowleri, indicating their critical roles in the survival and proliferation. This study emphasizes the significance of actin-binding proteins in understanding the infection and pathogenic mechanisms of N. fowleri.