Mutational analysis of a gene required for flagellar motility in the African Sleeping Sickness parasite
Trypanosoma brucei is a flagellated, unicellular protozoan that causes African Sleeping Sickness, a disease infecting humans and livestock in sub-Saharan Africa. The flagellum plays an integral role in pathogenicity of the organism. It is comprised of elements conserved in many organisms, making T. brucei a popular model for understanding flagellar components. Components of motile flagella (CMF) genes are found in all organisms with motile flagella. The knock down of CMF63 expression demonstrated that loss of the gene has moderate effects on flagellar motility. The gene has an EF-hand region, indicating that this protein may play a role in intracellular signaling. The purpose of my research is to better understand the function of CMF63 in T. brucei. Through homology searches, secondary structure, and helical wheel predictions, it was determined that a predicted α-helix from 393 404 amino acids may be involved in interactions with other secondary structures within the CMF63 protein or with other proteins. T. brucei was transfected with three different mutant CMF63 genes that were generated to perturb the potential interactions. By viewing the phenotypes of the mutants, it may be possible to ascertain if the α-helix plays an integral role in the function of CMF63 by interacting with another body.