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OPA1 deficiency causes mitochondrial fragmentation and also disrupts cristae, respiration, mitochondrial DNA (mtDNA) maintenance, and cell viability. It has not yet been established whether phenotypic severity can be modulated by genetic modifiers of OPA1. We screened the entire known mitochondrial proteome (1,531 genes) to identify genes that control mitochondrial morphology using a first\u2010in\u2010kind imaging pipeline. We identified 145 known and novel candidate genes whose depletion promoted elongation or fragmentation of the mitochondrial network in control fibroblasts and 91 in DOA+ patient fibroblasts that prevented mitochondrial fragmentation, including phosphatidyl glycerophosphate synthase (\n                    <jats:italic>PGS1<\/jats:italic>\n                    ). PGS1 depletion reduces CL content in mitochondria and rebalances mitochondrial dynamics in OPA1\u2010deficient fibroblasts by inhibiting mitochondrial fission, which improves defective respiration, but does not rescue mtDNA depletion, cristae dysmorphology, or apoptotic sensitivity. Our data reveal that the multifaceted roles of OPA1 in mitochondria can be functionally uncoupled by modulating mitochondrial lipid metabolism, providing novel insights into the cellular relevance of mitochondrial fragmentation.\n                  <\/jats:p>","DOI":"10.15252\/emmm.202013579","type":"journal-article","created":{"date-parts":[[2021,5,20]],"date-time":"2021-05-20T06:04:20Z","timestamp":1621490660000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":68,"title":["High\u2010throughput screening identifies suppressors of mitochondrial fragmentation in OPA1 fibroblasts"],"prefix":"10.1038","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8423-233X","authenticated-orcid":false,"given":"Emma","family":"Cretin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Priscilla","family":"Lopes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elodie","family":"Vimont","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0003-0587","authenticated-orcid":false,"given":"Takashi","family":"Tatsuta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1250-1462","authenticated-orcid":false,"given":"Thomas","family":"Langer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2922-3625","authenticated-orcid":false,"given":"Anastasia","family":"Gazi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5981-9166","authenticated-orcid":false,"given":"Martin","family":"Sachse","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7847-9320","authenticated-orcid":false,"given":"Patrick","family":"Yu\u2010Wai\u2010Man","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0802-4608","authenticated-orcid":false,"given":"Pascal","family":"Reynier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6770-6222","authenticated-orcid":false,"given":"Timothy","family":"Wai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,5,20]]},"reference":[{"key":"BFEMMM202013579_CR1","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1083\/jcb.201602007","volume":"213","author":"MJ Aaltonen","year":"2016","unstructured":"Aaltonen MJ, Friedman JR, Osman C, Salin B, di Rago J\u2010P, Nunnari J, Langer T, Tatsuta T (2016) MICOS and phospholipid transfer by Ups2\u2013Mdm35 organize membrane lipid synthesis in mitochondria. 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