Genome wide transcriptional change by oligomycin or manipulation of mitochondrial membrane potential
Oligomycin, an inhibitor of ATP synthase (Complex V), increases mitochondrial membrane potential (??m) and induces ISR. Since direct manipulation of mitochondrial membrane potential using the genetic reagent that we developed (UCP1) rescued the effect of oligomycin on ISR, we performed RNAseq to investigate the effect of direct manipulation of ??m by UCP1 on transcription. RNAseq analysis showed that direct manipulation of ??m does not induce a major transcriptional program. However, oligomycin treatment caused a significant change in the expression of many genes, and the activation of UCP1 under oligomycin treatment rescued most of the transcriptional changes. Overall design: C2C12 cells were infected with lentivirus carrying doxycycline-inducible UCP1. After 24 hr of seeding, Dox or water was added to induce protein expression. After 24 hr, various treatments (51 uM BSA, 300 uM oleic acid, 1 uM oligomycin + 300 uM oleic acid)were started in the presence of Dox or same volume of water. RNA was extracted 16 hr after the treatment and analyzed by RNA-sequencing.