{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:23:59Z","timestamp":1785425039257,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,22]]},"group-title":"Biochemistry","reference-count":74,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T00:00:00Z","timestamp":1784678400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R01GM118509"],"award-info":[{"award-number":["R01GM118509"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"award":["T32GM136534"],"award-info":[{"award-number":["T32GM136534"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005202","name":"Muscular Dystrophy Association","doi-asserted-by":"crossref","award":["1053188"],"award-info":[{"award-number":["1053188"]}],"id":[{"id":"10.13039\/100005202","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Bi-allelic mutations in\n                  <jats:italic>MSTO1<\/jats:italic>\n                  are linked to clinical disease phenotypes characteristic of mitochondrial dysfunction, including ataxia and muscular dystrophy. Consistent with this, MSTO1 patient-derived fibroblasts have fragmented mitochondria and a striking loss of mtDNA. Although MSTO1 has been implicated in regulating mitochondrial fusion, the molecular function of this cytosolic protein in vertebrate cells remains unclear. Using the auxin-inducible degradation (AID) system we demonstrate that MSTO1-FLAG-AID protein is rapidly depleted to almost undetectable levels. Importantly, these cells recapitulate the fragmented mitochondrial phenotype observed in patients and thus are a valuable model of disease. Surprisingly, prior to any changes in mitochondria, we show that MSTO1-depleted cells have a significant decrease in TRiC levels, an essential cytosolic ATP-dependent chaperone required to fold diverse substrates, including actin and tubulin. We reveal that TRiC is also reduced in MSTO1 patient-derived fibroblasts, indicating that loss of TRiC may contribute to disease pathophysiology. We further demonstrate that knockdown of TRiC leads to a decrease in MSTO1 protein levels and remarkably, was sufficient to induce a fragmented mitochondrial phenotype, independent of changes in tubulin or actin. This reveals a previously unrecognized connection between TRiC and mitochondrial homeostasis. Using co-immunoprecipitation we found that MSTO1 interacts with the TRiC chaperone. We also observe accumulation of early TRiC assembly subcomplexes in the absence of MSTO1 suggesting that MSTO1 facilitates assembly of TRiC. Together, our findings identify MSTO1 as a TRiC assembly factor and connect mitochondrial defects caused by MSTO1-depletion to the loss of TRiC.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance<\/jats:title>\n                  <jats:p>MSTO1, a protein linked to myopathy and ataxia, has been thought to control mitochondrial fusion, although the molecular mechanism is unknown. Using rapid depletion of MSTO1, we found that mitochondrial fragmentation appears only after six days. Significantly, the levels of the essential cytosolic chaperonin TRiC are reduced within two days of MSTO1 depletion. Directly depleting TRiC reproduces the fragmented mitochondrial phenotype seen with loss of MSTO1, consistent with a model where mitochondrial dysfunction is a downstream consequence of impaired protein folding rather than a direct effect of MSTO1 loss. We show that MSTO1 is required for assembly of TRiC, identifying it as a long-sought assembly factor for this macromolecular protein complex.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.21.739937","type":"posted-content","created":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T21:45:13Z","timestamp":1784756713000},"source":"Crossref","is-referenced-by-count":0,"title":["MSTO1 functions as a TRiC assembly factor linking cytosolic proteostasis to mitochondrial function"],"prefix":"10.64898","author":[{"given":"Amy M.","family":"Bounds","sequence":"first","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98195"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chloe","family":"Higuchi","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98195"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8070-3560","authenticated-orcid":false,"given":"Suzanne","family":"Hoppins","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98195"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026072705151903000_2026.07.21.739937v2.1","doi-asserted-by":"crossref","first-page":"398","DOI":"10.3390\/genes8120398","article-title":"The Mitochondrial Basis of Aging and Age-Related Disorders","volume":"8","year":"2017","journal-title":"Genes"},{"key":"2026072705151903000_2026.07.21.739937v2.2","doi-asserted-by":"publisher","DOI":"10.1111\/bph.15068"},{"key":"2026072705151903000_2026.07.21.739937v2.3","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2013.00292"},{"key":"2026072705151903000_2026.07.21.739937v2.4","doi-asserted-by":"publisher","DOI":"10.15252\/emmm.201607058"},{"key":"2026072705151903000_2026.07.21.739937v2.5","doi-asserted-by":"publisher","DOI":"10.1038\/s10038-020-00897-4"},{"key":"2026072705151903000_2026.07.21.739937v2.6","doi-asserted-by":"publisher","DOI":"10.1002\/humu.23262"},{"key":"2026072705151903000_2026.07.21.739937v2.7","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1016\/j.nmd.2019.03.011","article-title":"A novel case of MSTO1 gene related congenital muscular dystrophy with progressive neurological involvement","volume":"29","year":"2019","journal-title":"Neuromuscul. 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