Chromophore-Assisted Light Inactivation of Mitochondrial Electron Transport Chain Complex II in Caenorhabditis elegans
| Autor: | Alicia Y. Wei, Teresa A. Sherman, Thomas H. Foster, Andrew P. Wojtovich, Keith Nehrke |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Mitochondrial ROS Mutant Mutation Missense Respiratory chain Biology Mitochondrion 7. Clean energy Article 03 medical and health sciences Animals Respiratory function Caenorhabditis elegans Caenorhabditis elegans Proteins chemistry.chemical_classification Reactive oxygen species Multidisciplinary Electron Transport Complex II Genetic Complementation Test Clutch Size biology.organism_classification Mitochondria Optogenetics Chromophore-Assisted Light Inactivation 030104 developmental biology Biochemistry chemistry Gene Knockdown Techniques Larva |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep29695 |
| Popis: | Mitochondria play critical roles in meeting cellular energy demand, in cell death, and in reactive oxygen species (ROS) and stress signaling. Most Caenorhabditis elegans loss-of-function (lf) mutants in nuclear-encoded components of the respiratory chain are non-viable, emphasizing the importance of respiratory function. Chromophore-Assisted Light Inactivation (CALI) using genetically-encoded photosensitizers provides an opportunity to determine how individual respiratory chain components contribute to physiology following acute lf. As proof-of-concept, we expressed the ‘singlet oxygen generator’ miniSOG as a fusion with the SDHC subunit of respiratory complex II, encoded by mev-1 in C. elegans, using Mos1-mediated Single Copy Insertion. The resulting mev-1::miniSOG transgene complemented mev-1 mutant phenotypes in kn1 missense and tm1081(lf) deletion mutants. Complex II activity was inactivated by blue light in mitochondria from strains expressing active miniSOG fusions, but not those from inactive fusions. Moreover, light-inducible phenotypes in vivo demonstrated that complex II activity is important under conditions of high energy demand, and that specific cell types are uniquely susceptible to loss of complex II. In conclusion, miniSOG-mediated CALI is a novel genetic platform for acute inactivation of respiratory chain components. Spatio-temporally controlled ROS generation will expand our understanding of how the respiratory chain and mitochondrial ROS influence whole organism physiology. |
| Databáze: | OpenAIRE |
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