{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T04:55:34Z","timestamp":1789966534432,"version":"4.0.1"},"reference-count":35,"publisher":"The Company of Biologists","issue":"11","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2009,6,1]]},"abstract":"<jats:p>Clathrin-dependent endocytosis is believed to be involved in TGF\u03b2-stimulated cellular responses, but the subcellular locus at which TGF\u03b2 induces signaling remains unclear. Here, we demonstrate that inhibitors of clathrin-dependent endocytosis, which are known to arrest the progression of endocytosis at coated-pit stages, inhibit internalization of cell-surface-bound TGF\u03b2 and promote colocalization and accumulation of T\u03b2R-I and SARA at the plasma membrane. These inhibitors enhance TGF\u03b2-induced signaling and cellular responses (Smad2 phosphorylation\/nuclear localization and expression of PAI-1). Dynasore, a newly identified inhibitor of dynamin GTPase activity, is one of the most potent inhibitors among those tested and, furthermore, is a potent enhancer of TGF\u03b2. Dynasore ameliorates atherosclerosis in the aortic endothelium of hypercholesterolemic ApoE-null mice by counteracting the suppressed TGF\u03b2 responsiveness caused by the hypercholesterolemia, presumably acting through its effect on TGF\u03b2 endocytosis and signaling in vascular cells.<\/jats:p>","DOI":"10.1242\/jcs.038729","type":"journal-article","created":{"date-parts":[[2009,5,20]],"date-time":"2009-05-20T17:38:05Z","timestamp":1242841085000},"page":"1863-1871","source":"Crossref","is-referenced-by-count":123,"title":["Inhibitors of clathrin-dependent endocytosis enhance TGF\u03b2 signaling and responses"],"prefix":"10.1242","volume":"122","author":[{"given":"Chun-Lin","family":"Chen","sequence":"first","affiliation":[{"name":"Departments of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Doisy Research Center, St Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei-Hsien","family":"Hou","sequence":"additional","affiliation":[{"name":"Departments of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Doisy Research Center, St Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"I-Hua","family":"Liu","sequence":"additional","affiliation":[{"name":"Departments of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Doisy Research Center, St Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Hsiao","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Taipei Medical University, Taipei, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuan Shian","family":"Huang","sequence":"additional","affiliation":[{"name":"Auxagen, St Louis, MO 63132, USA"},{"name":"Center for Biotechnology and Biomedical Engineering, National Central University, Jhongli, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jung San","family":"Huang","sequence":"additional","affiliation":[{"name":"Departments of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Doisy Research Center, St Louis, MO 63104, USA"},{"name":"Institute of Systems Biology and Bioinformatics, National Central University, Jhongli, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","reference":[{"key":"2021042523544272700_REF1","doi-asserted-by":"crossref","unstructured":"Boensch, C., Huang, S. S., Connolly, D. T. and Huang, J. S. (1999). Cell surface retention sequence binding protein-1 interacts with the v-sis gene product and platelet-derived growth factor beta-type receptor in simian sarcoma virus-transformed cells. J. Biol. Chem.274, 10582-10589.","DOI":"10.1074\/jbc.274.15.10582"},{"key":"2021042523544272700_REF2","doi-asserted-by":"crossref","unstructured":"Bourdillon, M. C., Randon, J., Barek, L., Zibara, K., Chantal, C., Poston, R. N., Chignier, E. and McGregor, J. L. (2006). Reduced atherosclerotic lesion size in P-selectin deficient apolipoprotein E-knockout mice fed a chow but not a fat diet. J. Biomed. Biotechnol.2006, 49193.","DOI":"10.1155\/JBB\/2006\/49193"},{"key":"2021042523544272700_REF3","doi-asserted-by":"crossref","unstructured":"Chen, C. L., Huang, S. S. and Huang, J. S. (2006). Cellular heparan sulfate negatively modulates transforming growth factor-\u03b2 responsiveness in epithelial cells. J. Biol. Chem.281, 11506-11514.","DOI":"10.1074\/jbc.M512821200"},{"key":"2021042523544272700_REF4","doi-asserted-by":"crossref","unstructured":"Chen, C. L., Huang, S. S. and Huang, J. S. (2007). Cholesterol suppresses cellular TGF-\u03b2 responsiveness: implications in atherogenesis. J. Cell Sci.120, 3509-3521.","DOI":"10.1242\/jcs.006916"},{"key":"2021042523544272700_REF5","doi-asserted-by":"crossref","unstructured":"Chen, C. L., Huang, S. S. and Huang, J. S. (2008). Cholesterol modulates cellular TGF-\u03b2 responsiveness by altering TGF-\u03b2 binding to TGF-\u03b2 receptors. J. Cell. Physiol.215, 223-233.","DOI":"10.1002\/jcp.21303"},{"key":"2021042523544272700_REF6","doi-asserted-by":"crossref","unstructured":"Di Guglielmo, G. M., Le Roy, C., Goodfellow, A. F. and Wrana, J. L. (2003). Distinct endocytic pathways regulate TGF-\u03b2 receptor signalling and turnover. Nat. Cell Biol.5, 410-421.","DOI":"10.1038\/ncb975"},{"key":"2021042523544272700_REF7","doi-asserted-by":"crossref","unstructured":"Dickson, R. B., Willingham, M. C. and Pastan, I. H. (1982). Receptor-mediated endocytosis of alpha 2-macroglobulin: inhibition by ionophores and stimulation by Na+ and HCO3(-). Ann. NY Acad. Sci.401, 38-49.","DOI":"10.1111\/j.1749-6632.1982.tb25705.x"},{"key":"2021042523544272700_REF8","doi-asserted-by":"crossref","unstructured":"Dong, C., Li, Z., Alvarez, R., Jr., Feng, X. H. and Goldschmidt-Clermont, P. J. (2000). Microtubule binding to Smads may regulate TGF beta activity. Mol. Cell5, 27-34.","DOI":"10.1016\/S1097-2765(00)80400-1"},{"key":"2021042523544272700_REF9","doi-asserted-by":"crossref","unstructured":"Hansen, S. H., Sandvig, K. and van Deurs, B. (1993). Clathrin and HA2 adaptors: effects of potassium depletion, hypertonic medium, and cytosol acidification. J. Cell Biol.121, 61-72.","DOI":"10.1083\/jcb.121.1.61"},{"key":"2021042523544272700_REF10","doi-asserted-by":"crossref","unstructured":"Hayes, S., Chawla, A. and Corvera, S. (2002). TGF-\u03b2 receptor internalization into EEA1-enriched early endosomes: role in signaling to Smad2. J. Cell Biol.158, 1239-1249.","DOI":"10.1083\/jcb.200204088"},{"key":"2021042523544272700_REF11","doi-asserted-by":"crossref","unstructured":"Heldin, C. H., Miyazono, K. and ten Dijke, P. (1997). TGF-\u03b2 signalling from cell membrane to nucleus through SMAD proteins. Nature390, 465-471.","DOI":"10.1038\/37284"},{"key":"2021042523544272700_REF12","doi-asserted-by":"crossref","unstructured":"Horwitz, S. B., Chia, G. H., Harracksingh, C., Orlow, S., Pifko-Hirst, S., Schneck, J.,\n          Sorbara, L., Speaker, M., Wilk, E. W. and Rosen, O. M. (1981). Trifluoperazine inhibits phagocytosis in a macrophagelike cultured cell line. J. Cell Biol.91, 798-802.","DOI":"10.1083\/jcb.91.3.798"},{"key":"2021042523544272700_REF13","doi-asserted-by":"crossref","unstructured":"Huang, S. S. and Huang, J. S. (2005). TGF-\u03b2 control of cell proliferation. J. Cell. Biochem.96, 447-462.","DOI":"10.1002\/jcb.20558"},{"key":"2021042523544272700_REF14","doi-asserted-by":"crossref","unstructured":"Huang, S. S., Ling. T.-Y., Tang, F.-M., Yen, H.-Y., Leal, S. M. and Huang, J. S. (2003). Cellular growth inhibition by IGFBP-3 and TGF-\u03b21 requires LRP-1. FASEB J.17, 2068-2081.","DOI":"10.1096\/fj.03-0256com"},{"key":"2021042523544272700_REF15","doi-asserted-by":"crossref","unstructured":"Ito, T., Williams, J. D., Fraser, D. J. and Phillips, A. O. (2004). Hyaluronan regulates transforming growth factor-\u03b2 receptor compartmentalization. J. Biol. Chem.279, 25326-25332.","DOI":"10.1074\/jbc.M403135200"},{"key":"2021042523544272700_REF16","doi-asserted-by":"crossref","unstructured":"Kuratomi, Y., Akiyama, S., Ono, M., Shiraishi, N., Shimada, T., Ohkuma, S. and Kuwano, M. (1986). Thioridazine enhances lysosomal accumulation of epidermal growth factor and toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin. Exp. Cell Res.162, 436-448.","DOI":"10.1016\/0014-4827(86)90348-4"},{"key":"2021042523544272700_REF17","doi-asserted-by":"crossref","unstructured":"Le Roy, C. and Wrana, J. L. (2005). Clathrin- and non-clathrin-mediated endocytic regulation of cell signalling. Nat. Rev. Mol. Cell. Biol.6, 112-126.","DOI":"10.1038\/nrm1571"},{"key":"2021042523544272700_REF18","doi-asserted-by":"crossref","unstructured":"Li, M. O. and Flavell, R. A. (2008). Contextual regulation of inflammation: a duet by transforming growth factor-\u03b2 and interleukin-10. Immunity28, 468-476.","DOI":"10.1016\/j.immuni.2008.03.003"},{"key":"2021042523544272700_REF19","doi-asserted-by":"crossref","unstructured":"Ling, T. Y., Chen, C.-L., Huang, S. S. and Huang, J. S. (2004). Identification and characterization of the acidic pH binding sites for growth regulatory ligands of low density lipoprotein receptor-related protein-1. J. Biol. Chem.279, 38736-38748.","DOI":"10.1074\/jbc.M310537200"},{"key":"2021042523544272700_REF20","doi-asserted-by":"crossref","unstructured":"Lu, Z., Murray, J. T., Luo, W., Li, H., Wu, X., Xu, H., Backer, J. M. and Chen, Y. G. (2002). Transforming growth factor-\u03b2 activates Smad2 in the absence of receptor endocytosis. J. Biol. Chem.277, 29363-29368.","DOI":"10.1074\/jbc.M203495200"},{"key":"2021042523544272700_REF21","doi-asserted-by":"crossref","unstructured":"Macia, E., Ehrlich, M., Massol, R., Boucrot, E., Brunner, C. and Kirchhausen, T. (2006). Dynasore, a cell-permeable inhibitor of dynamin. Dev. Cell10, 839-850.","DOI":"10.1016\/j.devcel.2006.04.002"},{"key":"2021042523544272700_REF22","doi-asserted-by":"crossref","unstructured":"Massague, J. (1998). TGF-\u03b2 signal transduction. Annu. Rev. Biochem.67, 753-791.","DOI":"10.1146\/annurev.biochem.67.1.753"},{"key":"2021042523544272700_REF23","doi-asserted-by":"crossref","unstructured":"Metcalfe, J. C. and Grainger, D. J. (1995). Transforming growth factor-\u03b2 and the protection from cardiovascular injury hypothesis. Biochem. Soc. Trans.23, 403-406.","DOI":"10.1042\/bst0230403"},{"key":"2021042523544272700_REF24","doi-asserted-by":"crossref","unstructured":"Mitchell, H., Choudhury, A., Pagano, R. E. and Leof, E. B. (2004). Ligand-dependent and -independent transforming growth factor-\u03b2 receptor recycling regulated by clathrin-mediated endocytosis and Rab11. Mol. Biol. Cell15, 4166-4178.","DOI":"10.1091\/mbc.e04-03-0245"},{"key":"2021042523544272700_REF25","doi-asserted-by":"crossref","unstructured":"Mousavi, S. A., Malerod, L., Berg, T. and Kjeken, R. (2004). Clathrin-dependent endocytosis. Biochem. J.377, 1-16.","DOI":"10.1042\/bj20031000"},{"key":"2021042523544272700_REF26","doi-asserted-by":"crossref","unstructured":"Nankoe, S. R. and Sever, S. (2006). Dynasore puts a new spin on dynamin: a surprising dual role during vesicle formation. Trends Cell Biol.16, 607-609.","DOI":"10.1016\/j.tcb.2006.10.004"},{"key":"2021042523544272700_REF27","doi-asserted-by":"crossref","unstructured":"Penheiter, S. G., Mitchell, H., Garamszegi, N., Edens, M., Dore\u0301, J. J., Jr and Leof, E. B. (2002). Internalization-dependent and -independent requirements for transforming growth factor-\u03b2 receptor signaling via the Smad pathway. Mol. Cell. Biol.22, 4750-4759.","DOI":"10.1128\/MCB.22.13.4750-4759.2002"},{"key":"2021042523544272700_REF28","doi-asserted-by":"crossref","unstructured":"Roberts, A. B. (1998). Molecular and cell biology of TGF-\u03b2. Miner. Electrolyte Metab.24, 111-119.","DOI":"10.1159\/000057358"},{"key":"2021042523544272700_REF29","doi-asserted-by":"crossref","unstructured":"Rodal, S. K., Skretting, G., Garred, O., Vilhardt, F., van Deurs, B. and Sandvig, K. (1999). Extraction of cholesterol with methyl-beta-cyclodextrin perturbs formation of clathrin-coated endocytic vesicles. Mol. Biol. Cell10, 961-974.","DOI":"10.1091\/mbc.10.4.961"},{"key":"2021042523544272700_REF30","doi-asserted-by":"crossref","unstructured":"Salisbury, J. L., Condeelis, J. S. and Satir, P. (1980). Role of coated vesicles, microfilaments, and calmodulin in receptor-mediated endocytosis by cultured B lymphoblastoid cells. J. Cell Biol.87, 132-141.","DOI":"10.1083\/jcb.87.1.132"},{"key":"2021042523544272700_REF31","doi-asserted-by":"crossref","unstructured":"Schlegel, R., Dickson, R. B., Willingham, M. C. and Pastan, I. H. (1982). Amantadine and dansylcadaverine inhibit vesicular stomatitis virus uptake and receptor-mediated endocytosis of alpha 2-macroglobulin. Proc. Natl. Acad. Sci. USA79, 2291-2295.","DOI":"10.1073\/pnas.79.7.2291"},{"key":"2021042523544272700_REF32","doi-asserted-by":"crossref","unstructured":"Tsukazaki, T., Chiang, T. A., Davison, A. F., Attisano, L. and Wrana, J. L. (1998). SARA, a FYVE domain protein that recruits Smad2 to the TGF-\u03b2 receptor. Cell95, 779-791.","DOI":"10.1016\/S0092-8674(00)81701-8"},{"key":"2021042523544272700_REF33","doi-asserted-by":"crossref","unstructured":"Wang, L. H., Rothberg, K. G. and Anderson, R. G. (1993). Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J. Cell Biol.123, 1107-1117.","DOI":"10.1083\/jcb.123.5.1107"},{"key":"2021042523544272700_REF34","doi-asserted-by":"crossref","unstructured":"Xu, L., Chen, Y. G. and Massague, J. (2000). The nuclear import function of Smad2 is masked by SARA and unmasked by TGF-\u03b2-dependent phosphorylation. Nat. Cell Biol.2, 559-562.","DOI":"10.1038\/35019649"},{"key":"2021042523544272700_REF35","doi-asserted-by":"crossref","unstructured":"Zwaagstra, J. C., El-Alfy, M. and O'Connor-McCourt, M. D. (2001). Transforming growth factor (TGF)-\u03b2 1 internalization: modulation by ligand interaction with TGF-\u03b2 receptors types I and II and a mechanism that is distinct from clathrin-mediated endocytosis. J. Biol. Chem.276, 27237-27245.","DOI":"10.1074\/jbc.M100033200"}],"container-title":["Journal of Cell Science"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/journals.biologists.com\/jcs\/article-pdf\/122\/11\/1863\/1500917\/1863.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/journals.biologists.com\/jcs\/article-pdf\/122\/11\/1863\/1500917\/1863.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,4,26]],"date-time":"2021-04-26T00:21:08Z","timestamp":1619396468000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/jcs\/article\/122\/11\/1863\/35365\/Inhibitors-of-clathrin-dependent-endocytosis"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,6,1]]},"references-count":35,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2009,6,1]]}},"URL":"https:\/\/doi.org\/10.1242\/jcs.038729","relation":{},"ISSN":["1477-9137","0021-9533"],"issn-type":[{"value":"1477-9137","type":"electronic"},{"value":"0021-9533","type":"print"}],"subject":[],"published":{"date-parts":[[2009,6,1]]}}}