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Conjugation of ATG8s to single membranes (CASM) is a noncanonical LC3\u2011lipidation pathway activated by various stressors, distinct from canonical autophagy. We previously showed that TBT reduces lysosomal acidity and inhibits autophagy in SH-SY5Y cells. Furthermore, we observed TBT-induced LC3-II accumulation, which was reduced by bafilomycin A\n                    <jats:sub>1<\/jats:sub>\n                    , and tubular LC3-positive structures as hallmarks of CASM. In this study, we investigated whether TBT activates CASM. TBT (700\u00a0nM) induced LC3-II accumulation, which was completely blocked by bafilomycin A\n                    <jats:sub>1<\/jats:sub>\n                    in SH-SY5Y and HeLa cells. Unlike autophagy, TBT induced LC3-II accumulation even under class III PI3K inhibition by wortmannin and in FIP200-knockout cells. Salmonella effector protein SopF, which inhibits V-ATPase\n                    <jats:bold>\u2013<\/jats:bold>\n                    ATG16L1 association required for CASM, inhibited TBT-induced LC3-II accumulation. In FIP200-knockout cells, TBT induced LC3 accumulation on lysosomes, the primary CASM target. TBT also promoted nuclear translocation of transcription factor EB (TFEB) in a SopF-sensitive manner. Together, these results identify CASM as a lysosomal stress response to TBT, induced via the V-ATPase\n                    <jats:bold>\u2013<\/jats:bold>\n                    ATG16L1 axis, leading to TFEB activation. This mechanism provides a toxicological framework for understanding xenobiotic-induced lysosomal adaptations.\n                  <\/jats:p>","DOI":"10.1007\/s00204-026-04300-7","type":"journal-article","created":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T19:53:48Z","timestamp":1770494028000},"page":"1941-1951","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Tributyltin induces conjugation of ATG8s to single membranes via the V-ATPase\u2013ATG16L1 axis, leading to transcription factor EB activation in human cell lines"],"prefix":"10.1007","volume":"100","author":[{"given":"Shunichi","family":"Hatamiya","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6132-8286","authenticated-orcid":false,"given":"Masatsugu","family":"Miyara","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nanako","family":"Takahashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3461-1259","authenticated-orcid":false,"given":"Ami","family":"Oguro","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2645-3078","authenticated-orcid":false,"given":"Yaichiro","family":"Kotake","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,2,7]]},"reference":[{"issue":"5","key":"4300_CR1","doi-asserted-by":"publisher","first-page":"909","DOI":"10.1016\/j.molcel.2019.03.033","volume":"74","author":"H Chino","year":"2019","unstructured":"Chino H, Hatta T, Natsume T, Mizushima N (2019) Intrinsically disordered protein TEX264 mediates ER-phagy. 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