Chromophore-Assisted Light Inactivation of Mitochondrial Electron Transport Chain Complex II in Caenorhabditis elegans.

摘要: Mitochondria play critical roles in meeting cellular energy demand, cell death, and reactive oxygen species (ROS) stress signaling. Most Caenorhabditis elegans loss-of-function (lf) mutants nuclear-encoded components of the respiratory chain are non-viable, emphasizing importance function. Chromophore-Assisted Light Inactivation (CALI) using genetically-encoded photosensitizers provides an opportunity to determine how individual contribute physiology following acute lf. As proof-of-concept, we expressed ‘singlet generator’ miniSOG as a fusion with SDHC subunit complex II, encoded by mev-1 C. elegans, Mos1-mediated Single Copy Insertion. The resulting mev-1::miniSOG transgene complemented mutant phenotypes kn1 missense tm1081(lf) deletion mutants. Complex II activity was inactivated blue light mitochondria from strains expressing active fusions, but not those inactive fusions. Moreover, light-inducible vivo demonstrated that is important under conditions high specific types uniquely susceptible loss II. In conclusion, miniSOG-mediated CALI novel genetic platform for inactivation components. Spatio-temporally controlled ROS generation will expand our understanding mitochondrial influence whole organism physiology.