OPA1 ‐ dependent cristae modulation is essential for cellular adaptation to metabolic demand by David A Patten, Jacob Wong, Mireille Khacho, Vincent Soubannier, Ryan J Mailloux, Karine Pilon ‐ Larose, Jason G MacLaurin, David S Park, Heidi M McBride, Laura Trinkle ‐ Mulcahy, Mary ‐ Ellen Harper, Marc Germain, and Ruth S Slack EMBO J. Volume 33(22): November 18, 2014 ©2014 by European Molecular Biology Organization

Cristae condense, mitochondria elongate and OPA1 oligomerizes rapidly and reversibly during cell starvation MEFs were starved (STV) or not (Ctrl) for 2 h, fixed with 2% PFA and 1.6% glutaraldehyde and analysed by EM. Cristae and mitochondrial width were qu... David A Patten et al. EMBO J. 2014;33: ©2014 by European Molecular Biology Organization

OPA1 responds to energy substrate availability and responds by oligomerizing and maintaining cytochrome c in isolated mitochondria OPA1 oligomerization was analysed in isolated mouse liver mitochondria incubated with or without the indicated ETC substrate... David A Patten et al. EMBO J. 2014;33: ©2014 by European Molecular Biology Organization

OPA1 is required for resistance to starvation ‐ induced cell death independently of its fusion activity Cell death of WT and OPA1 KO cells starved or not for 6 h was analysed with propidium iodide (PI) and Hoechst (averages ± SEM of four independent experime... David A Patten et al. EMBO J. 2014;33: ©2014 by European Molecular Biology Organization

OPA1 regulates mitochondrial metabolism independently of mitochondrial fusion AWT and OPA1 knockout MEFs were infected with viruses encoding GFP, OPA1 or OPA1(Q297V) with dual promoters expressing GFP to allow for puromycin selection and FACS sorting cell... David A Patten et al. EMBO J. 2014;33: ©2014 by European Molecular Biology Organization

The dual role of OPA1 in regulating the ATP synthase AIsolated mitochondria incubated with complex I substrates were analysed by BN ‐ PAGE and blotted for ATP5A of the ATP synthase. David A Patten et al. EMBO J. 2014;33: ©2014 by European Molecular Biology Organization

OPA1 interacts with SLC25 proteins that regulate OPA1's response to starvation AMito ‐ YFP ‐ 3×flag, DIC ‐ 3×flag, OGC ‐ 3×flag, AGC1 ‐ 3×flag and AGC2 ‐ 3×flag constructs were transiently transfected into MEFs. Twenty ‐ four hours post ‐ transfection, cells were lysed,... David A Patten et al. EMBO J. 2014;33: ©2014 by European Molecular Biology Organization

Pharmacological blockade of SLC25A proteins drastically inhibit OPA1 oligomerization and function Isolated liver mitochondria were incubated with phenylsuccinate (PhS) or butylmalonate (BM), and OPA1 oligomerization was assessed as previously. David A Patten et al. EMBO J. 2014;33: ©2014 by European Molecular Biology Organization