{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T00:10:28Z","timestamp":1767831028921,"version":"3.49.0"},"reference-count":52,"publisher":"The Company of Biologists","issue":"4","license":[{"start":{"date-parts":[[1994,4,1]],"date-time":"1994-04-01T00:00:00Z","timestamp":765158400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/www.biologists.com\/user-licence-1-1\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[1994,4,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:p>The protein kinase inhibitor H-7 prevents the assembly of tight junctions in cultured Madin Darby Canine Kidney (MDCK) epithelial cells (Balda et al. (1991)J. Membr. Biol. 122, 193-202; Nigam et al. (1991)Biochem. Biophys. Res. Commun. 181, 548-553); however, its mechanism of action is unknown. To understand the basis of the activity of H-7 and other inhibitors we compared the effect of H-7 on the localization of proteins belonging to tight junctions and adherens-type junctions (zonula adhaerens and desmosome), and on the organization of actin microfilaments. Junction assembly was induced in MDCK cells either by the \u2018Ca2+ switch\u2019 procedure or by incubating trypsinized cells at normal extracellular Ca2+, and the cells were then immunofluorescently labeled with antibodies against cingulin, ZO-1, E-cadherin and desmoplakin, and with FITC-phalloidin. Here we show by measuring the transepithelial resistance that, in addition to H-7, H-8 and staurosporine can also significantly block the assembly of tight junctions, whereas HA1004 is poorly active. H-7 inhibited the accumulation of cingulin and ZO-1 in junctional areas most effectively when added during assembly at normal extracellular Ca2+. On the other hand, H-7 did not have major effects on the accumulation of E-cadherin and desmoplakin in the regions of cell-cell contact using either assembly protocol. Electron microscopy confirmed that H-7 does not abolish the formation of adherens-type junctions, suggesting that phosphorylation plays a different role in the assembly of tight junctions versus adherens-type junctions. Finally, in both protocols of junction assembly H-7 caused a major disorganization of actin microfilaments, suggesting that H-7 may prevent TJ assembly through its effect on the cytoskeleton.<\/jats:p>","DOI":"10.1242\/jcs.107.4.969","type":"journal-article","created":{"date-parts":[[2021,4,24]],"date-time":"2021-04-24T02:32:42Z","timestamp":1619231562000},"page":"969-981","source":"Crossref","is-referenced-by-count":40,"title":["Different effects of protein kinase inhibitors on the localization of junctional proteins at cell-cell contact sites"],"prefix":"10.1242","volume":"107","author":[{"given":"Natasha","family":"Denisenko","sequence":"first","affiliation":[{"name":"2 Department of Cell Biology, Cornell University Medical College, New York, NY 10021, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paolo","family":"Burighel","sequence":"additional","affiliation":[{"name":"1 Dipartimento di Biologia, Universita\u2019 di Padova, Via Trieste 75, Padova 35121, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sandra","family":"Citi","sequence":"additional","affiliation":[{"name":"1 Dipartimento di Biologia, Universita\u2019 di Padova, Via Trieste 75, Padova 35121, Italy"},{"name":"2 Department of Cell Biology, Cornell University Medical College, New York, NY 10021, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"237","published-online":{"date-parts":[[1994,4,1]]},"reference":[{"key":"2024022707090656100_JOCES_107_4_969C1","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1007\/BF01871420","article-title":"Assembly and sealing of tight junctions: possible participation of G-proteins, phospholipase C, protein kinase C and calmodulin","volume":"122","author":"Balda","year":"1991","journal-title":"J. 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