Characterizing TRAK & MIRO Interactions and localization to mitochondria
Lili Mitchell, Kathryn Reda, Claudia Vasquez, Hijab Fatima
Our research worked to characterize TRAK interactions with mitochondria that are MIRO-dependent and MIRO-independent. Using a similar approach to [Bocanegra et al. 2019], we used HeLa cells to quantify the interactions between MIRO and Trak proteins. As HeLa cells express both MIRO1 and MIRO2 on their own, we then switched our model system to mouse embryonic fibroblasts lacking MIRO expression. We could then control which MIRO protein the cells expressed, and in that way specifically investigate the TRAK interactions with either MIRO1 or MIRO2. We found that in the absence of MIRO, Trak still localized to mitochondria, indicating that TRAK has a MIRO-independent mitochondrial binding domain. When we examined the localization of Trak protein truncated to its MIRO-binding domain, the localization was diffuse in the cell. However, when we also expressed MIRO in those cells, the TRAK construct localized more strongly to mitochondria.
Biology, Biochemistry and Molecular Biology