Poster number 53 submitted by Vasilios Orologas
Super resolution nanoscopy of Trypanosoma brucei RNA binding protein 42 during acute infection in mice
Vasilios Orologas (Department of Microbiology, Biochemistry, and Molecular Genetics, Rutgers- New Jersey Medical School), Kezia Lizardo (Hackensack Meridian Health), Luke Fritzky (Rutgers- New Jersey Medical School ), Vivian Bellofatto (Department of Microbiology, Biochemistry, and Molecular Genetics, Rutgers- New Jersey Medical School)
RNA binding proteins (RBPs) are intricately involved in mRNA dynamics, including intracellular localization-based stability, and as reversible mediators of mRNA fate during cell stress. The complex life cycle, and polycistronic gene expression of Trypanosoma brucei, the African Trypanosome requires an extensive use of RBP-based regulatory machineries that aid parasite transition from insect vector to mammalian host. In the mammalian host, the parasite occupies and adapts to a variety of tissue microenvironments to generate peak parasitemia at day 6 post-infection. RBP42 is a polysome-associated, essential RNA binding protein, acting as a post-translational regulator of mRNA-encoding enzymes involved in T. brucei Central Carbon Metabolism. Our hypothesis is that the internal localization of RBP42 in T. brucei fluctuates to allow the parasites to derive nutrients from multiple host environments. To test this hypothesis, we are employing Leica Stellaris 8 tau-STED Super Resolution Microscopy, and experimental immunohistochemical staining to visualize the sub-cellular localization of RBP42 in parasites infecting lung, kidney, heart, pancreas, and adipose tissue in the mouse model. We will show Super Resolution images of parasites stained with anti-Variant Surface Glycoprotein antibodies (VSG), anti-RBP42 antibodies, and PicoGreen (nuclear and kinetoplast DNA stain) to reveal detailed images of parasites during acute infection.
Keywords: RNA binding proteins , STED microscopy , Trypanosoma brucei