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Schizophrenia is a chronic mental illness, characterized by severe symptoms such as delusions, hallucinations, disorganized speech, lack of motivation, and unpredictable behavior. The development of schizophrenia is caused by genetic and environmental factors. The complexity of these factors has made it difficult to develop new methods for the successful treatment of patients. Recent research has shown that disruption of proteostasis may also impact on the progression of schizophrenia. Disruption in proteostasis causes certain proteins to misfold and therefore aggregate if the cell fails to degrade them. Aggregation is a process in which large insoluble structures known as aggregates are formed. So far, five proteins have been identified that may aggregate in schizophrenia. Among these five is TRIO and F-actin-binding protein (TRIOBP-1), the protein of interest in this thesis. Using C-terminally truncated constructs, it was discovered that the critical region for aggregation of TRIOBP-1 is located in the central section of the protein. As a next step, different truncated variants of TRIOBP-1 were expressed in neuroblastoma cells and immunofluorescent microscopy was used as a way to visualize aggregation. In this way, the critical region for aggregation was narrowed down to a sequence of less than 10 amino acids. To expand and verify this, the next step was to use full-length constructs with a small deletion from their center. In this thesis, four such constructs were used. Unexpectedly, all four of them formed aggregates in cells. This finding indicates that either the PH domain, or an optional 59 disordered amino acids at the N-terminus have effect on TRIOBP-1’s aggregation propensity which was previously thought to be very unlikely. The next step is to generate plasmids lacking the optionally translated N-terminal unstructured region, and look into the effect it may have on aggregation. By generating a TRIOBP-1 mutant with the minimal number of mutations required to prevent aggregation, we will be able to generate model systems for studying TRIOBP-1 aggregation. This will allow us to better understand the role of TRIOBP-1 in the progression of schizophrenia. |
