Functional Characterization Of The Microtubule Severing Protein Katanin

Katanin is a member of the ATPase superfamily. It is composed of katanin60and katanin80subunits. Katanin60subunit has ATP hydrolysis activity and microtubule severing function. And katanin80subunit can locate katanin60in the centrosome. Currently, there is not a model animal for researching the function of katanin60except elegan. Therefore, the results in vitro have not been confirmed in vivo. Secondly, although studies in cell culture have clarified the katanin mutating could lead to the distruction of microtubule network. These studies are based on studies of cultured cells in vitro. Therefore it can not Completely explain the function of Katanin in the individual growth and development especially in the process of neuromuscular system development.We have bought the katanin60RNAi from the VDRC; using p factors local hop to get the mutant; connecting CDS of katanin60to pUAST vector, and micro injecting the vector into Drosophila’s embryos and then Filterring and positioning, Finally we got the stock overexpressed katanin60in Drosophila. Using Immunostaining and biochemical molecules to detect the protein expression and expression of position of overexpression, knockdown katanin60and mutant, and the phenotype in the neuromuscular system.The results show us that overexpression of katanin60resulted in MT fiber disrupted and a reduction in MT network especially in the perinuclear area and mutant can cause increased density of microtubules around the nucleus, while the performance was abnormal synaptic development, indicating that the synapses plays an important role in development.And in the cold treated recovery experiment, we see that the the exercise of functions needs a certain density of microtubule.