{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T13:45:57Z","timestamp":1789998357244,"version":"4.0.1"},"reference-count":42,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2018,12,7]],"date-time":"2018-12-07T00:00:00Z","timestamp":1544140800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["GB14- 36804G"],"award-info":[{"award-number":["GB14- 36804G"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"name":"UNCE - Univerzitn\u00ed v\u00fdzkumn\u00e1 centra, Charles University","award":["UNCE 204011"],"award-info":[{"award-number":["UNCE 204011"]}]},{"name":"PRVOUK - Programy rozvoje v\u011bdn\u00edch oblast\u00ed na Univerzit\u011b Karlov\u011b, Charles University","award":["P24\/LF1\/3"],"award-info":[{"award-number":["P24\/LF1\/3"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mitochondrial protein quality control is crucial for the maintenance of correct mitochondrial homeostasis. It is ensured by several specific mitochondrial proteases located across the various mitochondrial subcompartments. Here, we focused on characterization of functional overlap and cooperativity of proteolytic subunits AFG3L2 (AFG3 Like Matrix AAA Peptidase Subunit 2) and YME1L (YME1 like ATPase) of mitochondrial inner membrane AAA (ATPases Associated with diverse cellular Activities) complexes in the maintenance of mitochondrial structure and respiratory chain integrity. We demonstrate that loss of AFG3L2 and YME1L, both alone and in combination, results in diminished cell proliferation, fragmentation of mitochondrial reticulum, altered cristae morphogenesis, and defective respiratory chain biogenesis. The double AFG3L2\/YME1L knockdown cells showed marked upregulation of OPA1 protein forms, with the most prominent increase in short OPA1 (optic atrophy 1). Loss of either protease led to marked elevation in OMA1 (OMA1 zinc metallopeptidase) (60 kDa) and severe reduction in the SPG7 (paraplegin) subunit of the m-AAA complex. Loss of the YME1L subunit led to an increased Drp1 level in mitochondrial fractions. While loss of YME1L impaired biogenesis and function of complex I, knockdown of AFG3L2 mainly affected the assembly and function of complex IV. Our results suggest cooperative and partly redundant functions of AFG3L2 and YME1L in the maintenance of mitochondrial structure and respiratory chain biogenesis and stress the importance of correct proteostasis for mitochondrial integrity.<\/jats:p>","DOI":"10.3390\/ijms19123930","type":"journal-article","created":{"date-parts":[[2018,12,10]],"date-time":"2018-12-10T03:36:41Z","timestamp":1544413001000},"page":"3930","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":37,"title":["Loss of Mitochondrial AAA Proteases AFG3L2 and YME1L Impairs Mitochondrial Structure and Respiratory Chain Biogenesis"],"prefix":"10.3390","volume":"19","author":[{"given":"Jana","family":"Cesnekova","sequence":"first","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, 12808 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Rodinova","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, 12808 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hana","family":"Hansikova","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, 12808 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiri","family":"Zeman","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, 12808 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3739-7246","authenticated-orcid":false,"given":"Lukas","family":"Stiburek","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Adolescent Medicine, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, 12808 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,12,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"34","DOI":"10.3389\/fmolb.2017.00034","article-title":"Multifunctional Mitochondrial AAA Proteases","volume":"4","author":"Glynn","year":"2017","journal-title":"Front. Mol. Biosci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"R170","DOI":"10.1016\/j.cub.2018.01.004","article-title":"Mitophagy and Quality Control Mechanisms in Mitochondrial Maintenance","volume":"28","author":"Pickles","year":"2018","journal-title":"Curr. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1111\/boc.201600019","article-title":"Intersection of mitochondrial fission and fusion machinery with apoptotic pathways: Role of Mcl-1","volume":"108","author":"Morciano","year":"2016","journal-title":"Biol. Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"4737","DOI":"10.1021\/acs.biochem.7b00663","article-title":"Metalloproteases of the Inner Mitochondrial Membrane","volume":"56","author":"Levytskyy","year":"2017","journal-title":"Biochemistry"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1080\/10409230701380452","article-title":"Protein degradation within mitochondria: Versatile activities of AAA proteases and other peptidases","volume":"42","author":"Koppen","year":"2007","journal-title":"Crit. Rev. Biochem. Mol. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1042\/EBC20160009","article-title":"Protein quality control at the mitochondrion","volume":"60","author":"Voos","year":"2016","journal-title":"Essays Biochem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1091\/mbc.e11-08-0674","article-title":"YME1L controls the accumulation of respiratory chain subunits and is required for apoptotic resistance, cristae morphogenesis, and cell proliferation","volume":"23","author":"Stiburek","year":"2012","journal-title":"Mol. Biol. Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e896","DOI":"10.1038\/cddis.2013.414","article-title":"Loss of Yme1L perturbates mitochondrial dynamics","volume":"4","author":"Ruan","year":"2013","journal-title":"Cell Death Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"908","DOI":"10.1016\/j.cmet.2013.11.006","article-title":"Stress-regulated translational attenuation adapts mitochondrial protein import through Tim17A degradation","volume":"18","author":"Rainbolt","year":"2013","journal-title":"Cell Metab."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2041","DOI":"10.1016\/j.celrep.2016.02.011","article-title":"Reciprocal Degradation of YME1L and OMA1 Adapts Mitochondrial Proteolytic Activity during Stress","volume":"14","author":"Rainbolt","year":"2016","journal-title":"Cell Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1042\/BJ20151029","article-title":"The mammalian homologue of yeast Afg1 ATPase (lactation elevated 1) mediates degradation of nuclear-encoded complex IV subunits","volume":"473","author":"Cesnekova","year":"2016","journal-title":"Biochem. J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1038\/emboj.2011.477","article-title":"Nonsense mutations in the COX1 subunit impair the stability of respiratory chain complexes rather than their assembly","volume":"31","author":"Tatsuta","year":"2012","journal-title":"EMBO J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1083\/jcb.201504062","article-title":"Quality control of mitochondrial protein synthesis is required for membrane integrity and cell fitness","volume":"211","author":"Richter","year":"2015","journal-title":"J. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4388","DOI":"10.1073\/pnas.1702938114","article-title":"Proteolytic control of the mitochondrial calcium uniporter complex","volume":"114","author":"Tsai","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Consolato, F., Maltecca, F., Tulli, S., Sambri, I., and Casari, G. (2018). m-AAA and i-AAA complexes coordinate to regulate OMA1, the stress-activated supervisor of mitochondrial dynamics. J. Cell Sci., 131.","DOI":"10.1242\/jcs.213546"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.molcel.2015.08.009","article-title":"SPG7 Is an Essential and Conserved Component of the Mitochondrial Permeability Transition Pore","volume":"60","author":"Shanmughapriya","year":"2015","journal-title":"Mol. Cell"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Magri, S., Fracasso, V., Plumari, M., Alfei, E., Ghezzi, D., Gellera, C., Rusmini, P., Poletti, A., Bella, D.D., and Elia, A.E. (2018). Concurrent AFG3L2 and SPG7 mutations associated with syndromic parkinsonism and optic atrophy with aberrant OPA1 processing and mitochondrial network fragmentation. Hum. Mutat., 39.","DOI":"10.1002\/humu.23658"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Hartmann, B., Wai, T., Hu, H., MacVicar, T., Musante, L., Fischer-Zirnsak, B., Stenzel, W., Gr\u00e4f, R., van den Heuvel, L., and Ropers, H.H. (2016). Homozygous YME1L1 mutation causes mitochondriopathy with optic atrophy and mitochondrial network fragmentation. Elife, 5.","DOI":"10.7554\/eLife.16078"},{"key":"ref_19","unstructured":"Sprenger, H.G., Wani, G., Hesseling, A., K\u00f6nig, T., Patron, M., MacVicar, T., Ahola, S., Wai, T., Barth, E., and Rugarli, E.I. (2018). Loss of the mitochondrial i-AAA protease YME1L leads to ocular dysfunction and spinal axonopathy. EMBO Mol. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"47687","DOI":"10.18632\/oncotarget.9959","article-title":"LACE1 interacts with p53 and mediates its mitochondrial translocation and apoptosis","volume":"7","author":"Cesnekova","year":"2016","journal-title":"Oncotarget"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"S27","DOI":"10.33549\/physiolres.930000.55.S2.27","article-title":"Biogenesis of eukaryotic cytochrome c oxidase","volume":"55","author":"Stiburek","year":"2006","journal-title":"Physiol. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1002\/embj.201386474","article-title":"Stress-induced OMA1 activation and autocatalytic turnover regulate OPA1-dependent mitochondrial dynamics","volume":"33","author":"Baker","year":"2014","journal-title":"EMBO J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1083\/jcb.200906084","article-title":"Regulation of OPA1 processing and mitochondrial fusion by m-AAA protease isoenzymes and OMA1","volume":"187","author":"Ehses","year":"2009","journal-title":"J. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1038\/sj.emboj.7601963","article-title":"Fission and selective fusion govern mitochondrial segregation and elimination by autophagy","volume":"27","author":"Twig","year":"2008","journal-title":"EMBO J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2676","DOI":"10.15252\/embj.201488349","article-title":"OPA1-dependent cristae modulation is essential for cellular adaptation to metabolic demand","volume":"33","author":"Patten","year":"2014","journal-title":"EMBO J."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2557","DOI":"10.1016\/j.celrep.2017.05.073","article-title":"OPA1 Isoforms in the Hierarchical Organization of Mitochondrial Functions","volume":"19","author":"Mishra","year":"2017","journal-title":"Cell Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.cell.2006.06.025","article-title":"OPA1 controls apoptotic cristae remodeling independently from mitochondrial fusion","volume":"126","author":"Frezza","year":"2006","journal-title":"Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/j.cell.2006.09.021","article-title":"Mitochondrial inner-membrane fusion and crista maintenance requires the dynamin-related GTPase Mgm1","volume":"127","author":"Meeusen","year":"2006","journal-title":"Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1083\/jcb.201308006","article-title":"The i-AAA protease YME1L and OMA1 cleave OPA1 to balance mitochondrial fusion and fission","volume":"204","author":"Anand","year":"2014","journal-title":"J. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2297","DOI":"10.1242\/jcs.159186","article-title":"OPA1 processing in cell death and disease\u2014The long and short of it","volume":"129","author":"MacVicar","year":"2016","journal-title":"J. Cell Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1083\/jcb.200906083","article-title":"Inducible proteolytic inactivation of OPA1 mediated by the OMA1 protease in mammalian cells","volume":"187","author":"Head","year":"2009","journal-title":"J. Cell Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3858","DOI":"10.1093\/hmg\/dds214","article-title":"Respiratory dysfunction by AFG3L2 deficiency causes decreased mitochondrial calcium uptake via organellar network fragmentation","volume":"21","author":"Maltecca","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1161\/CIRCRESAHA.117.311143","article-title":"TNFR2 Stimulation Promotes Mitochondrial Fusion via Stat3- and NF-kB-Dependent Activation of OPA1 Expression","volume":"121","author":"Nan","year":"2017","journal-title":"Circ. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1002\/embr.201338240","article-title":"Membrane depolarization activates the mitochondrial protease OMA1 by stimulating self-cleavage","volume":"15","author":"Zhang","year":"2014","journal-title":"EMBO Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1111\/j.1749-6632.2010.05629.x","article-title":"Dynamic regulation of mitochondrial fission through modification of the dynamin-related protein Drp1","volume":"1201","author":"Chang","year":"2010","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4216","DOI":"10.1091\/mbc.e09-03-0218","article-title":"Autocatalytic processing of m-AAA protease subunits in mitochondria","volume":"20","author":"Koppen","year":"2009","journal-title":"Mol. Biol. Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1083\/jcb.200304112","article-title":"Loss of m-AAA protease in mitochondria causes complex I deficiency and increased sensitivity to oxidative stress in hereditary spastic paraplegia","volume":"163","author":"Atorino","year":"2003","journal-title":"J. Cell Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2827","DOI":"10.1523\/JNEUROSCI.4677-07.2008","article-title":"The mitochondrial protease AFG3L2 is essential for axonal development","volume":"28","author":"Maltecca","year":"2008","journal-title":"J. Neurosci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1083\/jcb.2.6.799","article-title":"Permanganate: A new fixative for electron microscopy","volume":"2","author":"Luft","year":"1956","journal-title":"J. Biophys. Biochem. Cytol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1016\/j.jmb.2007.09.044","article-title":"Knockdown of human Oxa1l impairs the biogenesis of F1Fo-ATP synthase and NADH:ubiquinone oxidoreductase","volume":"374","author":"Stiburek","year":"2007","journal-title":"J. Mol. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/978-1-62703-236-0_6","article-title":"Identification of p53 in mitochondria","volume":"962","author":"Vaseva","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1042\/BJ20050807","article-title":"Tissue-specific cytochrome c oxidase assembly defects due to mutations in SCO2 and SURF1","volume":"392","author":"Stiburek","year":"2005","journal-title":"Biochem. J."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/12\/3930\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:32:00Z","timestamp":1760196720000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/12\/3930"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,12,7]]},"references-count":42,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["ijms19123930"],"URL":"https:\/\/doi.org\/10.3390\/ijms19123930","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,12,7]]}}}