{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T21:25:19Z","timestamp":1768512319149,"version":"3.49.0"},"posted":{"date-parts":[[2024,5,30]]},"group-title":"Cell Biology","reference-count":51,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,5,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Myofibroblast differentiation, characterized by accumulation of cytoskeletal and extracellular matrix proteins by fibroblasts, is a key process in wound healing and pathogenesis of tissue fibrosis. Transforming growth factor-\u03b2 (TGF-\u03b2) is the most powerful known driver of myofibroblast differentiation. TGF-\u03b2 signals through transmembrane receptor serine\/threonine kinases that phosphorylate Smad transcription factors (Smad2\/3) leading to activation of transcription of target genes. Heterotrimeric G proteins mediate a distinct signaling from seven-transmembrane G protein coupled receptors, not commonly linked to Smad activation. We asked if G protein signaling plays any role in TGF-\u03b2-induced myofibroblast differentiation, using primary cultured human lung fibroblasts. Activation of G\u03b1s by cholera toxin blocked TGF-\u03b2-induced myofibroblast differentiation without affecting Smad2\/3 phosphorylation. Inhibition of G\u03b1i by pertussis toxin, or siRNA-mediated combined knockdown of G\u03b1q and G\u03b111 had no significant effect on TGF-\u03b2-induced myofibroblast differentiation. A combined knockdown of G\u03b112 and G\u03b113 resulted in a drastic inhibition of TGF-\u03b2-stimulated expression of myofibroblast marker proteins (collagen-1, fibronectin, smooth-muscle \u03b1-actin), with siG\u03b112 being significantly more potent than siG\u03b113. Mechanistically, a combined knockdown of G\u03b112 and G\u03b113 resulted in a substantially reduced phosphorylation of Smad2 and Smad3 in response to TGF-\u03b2, which was accompanied by a significant decrease in the expression of TGF\u03b2 receptors (TGFBR1, TGFBR2) and of Smad3 under siG\u03b112\/13 conditions. In conclusion, our study uncovers a novel role of G\u03b112\/13 proteins in the control of TGF-\u03b2 signaling and myofibroblast differentiation.<\/jats:p>","DOI":"10.1101\/2024.05.29.596473","type":"posted-content","created":{"date-parts":[[2024,5,30]],"date-time":"2024-05-30T16:10:15Z","timestamp":1717085415000},"source":"Crossref","is-referenced-by-count":0,"title":["Critical role of G\u03b112 and G\u03b113 proteins in TGF-\u03b2-induced myofibroblast differentiation"],"prefix":"10.64898","author":[{"given":"Eleanor B.","family":"Reed","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Albert","family":"Sitikov","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8909-356X","authenticated-orcid":false,"given":"Robert B.","family":"Hamanaka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Reng\u00fcl","family":"Cetin-Atalay","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2056-612X","authenticated-orcid":false,"given":"G\u00f6khan M.","family":"Mutlu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2501-3778","authenticated-orcid":false,"given":"Alexander A.","family":"Mongin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nickolai O.","family":"Dulin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024060120501170000_2024.05.29.596473v1.1","first-page":"4776","article-title":"Synergistic interaction of two classes of transforming growth factors from murine sarcoma cells","volume":"42","year":"1982","journal-title":"Cancer Res"},{"key":"2024060120501170000_2024.05.29.596473v1.2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.80.20.6264"},{"key":"2024060120501170000_2024.05.29.596473v1.3","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.1.119"},{"key":"2024060120501170000_2024.05.29.596473v1.4","doi-asserted-by":"crossref","unstructured":"Massague, J. and Sheppard, D. 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