caa a22 4500 606177361 CHVBK 20210128100751.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s00018-015-1863-9 doi (NATIONALLICENCE)springer-10.1007/s00018-015-1863-9 Mitochondrial fusion provides an ‘initial metabolic complementation' controlled by mtDNA [Elektronische Daten] [Liang Yang, Qi Long, Jinglei Liu, Haite Tang, Yuxing Li, Feixiang Bao, Dajiang Qin, Duanqing Pei, Xingguo Liu] Heteroplasmic cells, harboring both mutant and normal mitochondrial DNAs (mtDNAs), must accumulate mutations to a threshold level before respiratory activity is affected. This phenomenon has led to the hypothesis of mtDNA complementation by inter-mitochondrial content mixing. The precise mechanisms of heteroplasmic complementation are unknown, but it depends both on the mtDNA nucleoid dynamics among mitochondria as well as the mitochondrial dynamics as influenced by mtDNA. We tracked nucleoids among the mitochondria in real time to show that they are shared after complete fusion but not ‘kiss-and-run'. Employing a cell hybrid model, we further show that mtDNA-less mitochondria, which have little ATP production and extensive Opa1 proteolytic cleavage, exhibit weak fusion activity among themselves, yet remain competent in fusing with healthy mitochondria in a mitofusin- and OPA1-dependent manner, resulting in restoration of metabolic function. Depletion of mtDNA by overexpression of the matrix-targeted nuclease UL12.5 resulted in heterogeneous mitochondrial membrane potential (ΔΨm) at the organelle level in mitofusin-null cells but not in wild type. In this system, overexpression of mitofusins or application of the fusion-promoting drug M1 could partially rescue the metabolic damage caused by UL12.5. Interestingly, mtDNA transcription/translation is not required for normal mitochondria to restore metabolic function to mtDNA-less mitochondria by fusion. Thus, interplay between mtDNA and fusion capacity governs a novel ‘initial metabolic complementation'. Springer Basel, 2015 Tfam nationallicence Rho0 nationallicence Photoactivation nationallicence Mfn1 nationallicence Mfn2 nationallicence Nucleoids nationallicence Cap : Chloramphenicol nationallicence DMEM : Dulbecco's modified Eagle's medium nationallicence EB : Ethidium bromide nationallicence FBS : Fetal bovine serum nationallicence FI : Fluorescence intensity nationallicence IMM : Inner mitochondrial membrane nationallicence KFP : Kindling fluorescent protein nationallicence MEF : Mouse embryonic fibroblast nationallicence mt : Mitochondrial nationallicence mtFP : Mitochondrial matrix-targeting fluorescent protein nationallicence mtDNAs : Mitochondrial DNAs nationallicence OMM : Outer mitochondrial membrane nationallicence PAGFP : Photoactivatable green fluorescent protein nationallicence PBS : Phosphate-buffered saline nationallicence PEG : Polyethylene glycol nationallicence Q-PCR : Quantitative polymerase chain reaction. Rho0 cells: cells lacking mtDNA nationallicence Tfam : Mitochondrial transcription factor A nationallicence TMRM : Tetramethylrhodamine methyl ester nationallicence VK3 : Vitamin K3 nationallicence WT : Wild type nationallicence ΔΨm : Mitochondrial membrane potential nationallicence Yang Liang Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Long Qi Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Liu Jinglei Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Tang Haite Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Li Yuxing Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Bao Feixiang Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Qin Dajiang Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Pei Duanqing Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Liu Xingguo Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut Cellular and Molecular Life Sciences Springer Basel 72/13(2015-07-01), 2585-2598 1420-682X 72:13<2585 2015 72 18 https://doi.org/10.1007/s00018-015-1863-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1863-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yang Liang Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Long Qi Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Liu Jinglei Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Tang Haite Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Li Yuxing Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Bao Feixiang Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Qin Dajiang Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Pei Duanqing Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 700 1- Liu Xingguo Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530, Guangzhou, China aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/13(2015-07-01), 2585-2598 1420-682X 72:13<2585 2015 72 18