{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,17]],"date-time":"2026-05-17T07:54:30Z","timestamp":1779004470939,"version":"3.51.4"},"posted":{"date-parts":[[2023,10,14]]},"group-title":"Cell Biology","reference-count":81,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,10,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Non-alcoholic fatty liver disease (NAFLD), recently renamed metabolic dysfunction-associated steatotic liver disease (MASLD), is a progressive metabolic disorder that begins with aberrant triglyceride accumulation in the liver and can lead to cirrhosis and cancer. A common variant in the gene\n                  <jats:italic>PNPLA3<\/jats:italic>\n                  , encoding the protein PNPLA3-I148M, is the strongest known genetic risk factor for MASLD to date. Despite its discovery twenty years ago, the function of PNPLA3, and now the role of PNPLA3-I148M, remain unclear. In this study, we sought to dissect the biogenesis of PNPLA3 and PNPLA3-I148M and characterize changes induced by endogenous expression of the disease-causing variant. Contrary to bioinformatic predictions and prior studies with overexpressed proteins, we demonstrate here that PNPLA3 and PNPLA3-I148M are not endoplasmic reticulum-resident transmembrane proteins. To identify their intracellular associations, we generated a paired set of isogenic human hepatoma cells expressing PNPLA3 and PNPLA3-I148M at endogenous levels. Both proteins were enriched in lipid droplet, Golgi, and endosomal fractions. Purified PNPLA3 and PNPLA3-I148M proteins associated with phosphoinositides commonly found in these compartments. Despite a similar fractionation pattern as the wild-type variant, PNPLA3-I148M induced morphological changes in the Golgi apparatus, including increased lipid droplet-Golgi contact sites, which were also observed in I148M-expressing primary human patient hepatocytes. In addition to lipid droplet accumulation, PNPLA3-I148M expression caused significant proteomic and transcriptomic changes that resembled all stages of liver disease. Cumulatively, we validate an endogenous human cellular system for investigating PNPLA3-I148M biology and identify the Golgi apparatus as a central hub of PNPLA3-I148M-driven cellular change.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance Statement<\/jats:title>\n                  <jats:p>\n                    Fatty liver disease affects nearly a quarter of the world\u2019s population and has both environmental and genetic risk factors. A mutation in the gene\n                    <jats:italic>PNPLA3<\/jats:italic>\n                    that converts Ile 148 to Met is the strongest known genetic risk factor for developing fatty liver disease. Using a series of techniques to track endogenous PNPLA3 and PNPLA3-I148M biogenesis and localization, we reveal new insights into how the mutation changes cellular dynamics. Although previous reports focus on its role on lipid droplets, we reveal that PNPLA3-I148M also functions at the Golgi apparatus, an organelle critical for protein transport into and out of the cell and lipid signaling. PNPLA3-I148M causes altered Golgi morphology and drives changes reminiscent of liver disease.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.10.13.562302","type":"posted-content","created":{"date-parts":[[2023,10,14]],"date-time":"2023-10-14T15:55:12Z","timestamp":1697298912000},"source":"Crossref","is-referenced-by-count":4,"title":["The fatty liver disease-causing protein PNPLA3-I148M alters lipid droplet-Golgi dynamics"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2120-0971","authenticated-orcid":false,"given":"David J.","family":"Sherman","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lei","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer L.","family":"Mamrosh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiansong","family":"Xie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John","family":"Ferbas","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brett","family":"Lomenick","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark S.","family":"Ladinsky","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rati","family":"Verma","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ingrid C.","family":"Rulifson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raymond J.","family":"Deshaies","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023101902500929000_2023.10.13.562302v1.1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2019-318813"},{"key":"2023101902500929000_2023.10.13.562302v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(20)32511-3"},{"key":"2023101902500929000_2023.10.13.562302v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/ng.257"},{"key":"2023101902500929000_2023.10.13.562302v1.4","doi-asserted-by":"publisher","DOI":"10.1002\/hep.24283"},{"key":"2023101902500929000_2023.10.13.562302v1.5","doi-asserted-by":"crossref","first-page":"e14324","DOI":"10.1097\/MD.0000000000014324","article-title":"Association between PNPLA3 rs738409 polymorphism and nonalcoholic fatty liver disease (NAFLD) susceptibility and severity: A meta-analysis","volume":"98","year":"2019","journal-title":"Medicine (Baltimore"},{"key":"2023101902500929000_2023.10.13.562302v1.6","doi-asserted-by":"crossref","unstructured":"N. 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