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Fig. 8 ATP-binding and ATP-hydrolysis mutations in Torsin1 alter cellular localization.
MN9D cells were transfected with expression plasmids encoding either (A) Torsin1a-eGFP or (B) Torsin1b-eGFP fusion proteins. The images show single confocal planes through transfected cells demonstrating GFP fusion protein localization (green, left column) relative to the nucleus (DAPI counterstain, blue, center column). For each panel, the top row of images shows wild type Torsin1, the middle row of images shows a Torsin1[K114T] mutant, which disrupts the Walker A (ATP-binding) domain, and the bottom row shows a Torsin1[E177Q] mutant, which disrupts the Walker B (ATP hydrolysis) domain. (C) Four independent samples for each construct were evaluated for nuclear envelope localization of the fusion protein. The graphs show the mean % cells with nuclear envelope localization (100–200 cells were scored for each construct; error bars show standard error; ***P<0.0001, *p<0.05 versus WT, heteroscedastic 2-tailed T-test with Bonferroni correction).