Regulatory Role of TOR Enzyme on Cellular Metabolism and Morphological Transformation in Naegleria Amoeba

Abstract

Record ID: 259

Mentorship Award: Excellence in Research Mentoring

Type: Podium Presentation (in-person)

Advisor: Yoshi Odaka

Abstract: Naegleria fowleri is a pathogenic unicellular protist free living ameba commonly found in warm freshwater. Depending on the environment, it undergoes three stages of life cycle - trophozoite (infectious), flagellate (motile), cyst (dormant). The trophozoite form infects the central nervous system by entering the brain through the olfactory nerve in the nasal cavity, causing primary amoebic meningoencephalitis (PAM). The infection is rapid and fatal, with a mortality rate of 97%. Our central idea is to promote cyst formation in Naelgeria, followed by targeting drugs to treat PAM. In this study, regulatory role of target of rapamycin (TOR) kinase in Naegleria gruberi, a non-pathogenic species, was assessed. Phosphoproteomics analysis identified phosphorylation of glycolytic enzyme enolase and enolase-associating proteins when TOR was activated, making it an attractive candidate as a downstream target of TOR. Therefore, GFP-tagged Naegleria enolase was engineered and expressed in N. gruberi and murine MK4 cells, and subsequently, point mutations were introduced to the TOR-sensitive phosphorylation sites. We are currently investigating the effects of the mutations on enolase subcellular localization.