Unraveling Cellular Phenotypes of Novel TorsinA/TOR1A Mutations

ABSTRACT A three‐nucleotide (GAG) deletion (ΔE) in TorsinA (TOR1A) has been identified as the most common cause of dominantly inherited early‐onset torsion dystonia (DYT1). TOR1A encodes a chaperone‐like AAA+‐protein localized in the endoplasmic reticulum. Currently, only three additional, likely mutations have been reported in single dystonia patients. Here, we report two new, putative TOR1A mutations (p.A14_P15del and p.E121K) that we examined functionally in comparison with wild‐type (WT) protein and two known mutations (ΔE and p.R288Q). While inclusion formation is a characteristic feature for ΔE TOR1A, elevated levels of aggregates for other mutations were not observed when compared with WT TOR1A. WT and mutant TOR1A showed preferred degradation through the autophagy‐lysosome pathway, which is most pronounced for p.A14_P15del, p.R288Q, and ΔE TOR1A. Notably, blocking of the autophagy pathway with bafilomycin resulted in a significant increase in inclusion formation in p.E121K TOR1A. In addition, all variants had an influence on protein stability. Although the p.A14_P15del mutation affects the proposed oligomerization domain of TOR1A, this mutation did not disturb the ability to dimerize. Our findings demonstrate functional changes for all four mutations on different levels. Thus, both diagnostic and research genetic screening of dystonia patients should not be limited to testing for the ∆E mutation. We report two new putative TOR1A mutations (p.A14_P15del and p.E121K) that we examined functionally in comparison with wildtype protein. Our findings demonstrate functional changes on different levels.Western blot analysis of cross‐linking experiments with formaldehyde (FA) and quantification of TOR1A levels in SH‐SY5Y cells expressing TOR1A show significantly enhanced dimerization at 74 kDa for p.A14_P15del mutant TOR1A (NT: non‐treated).