Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia

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Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia

BR Stevenson, JD Siliciano, MS Mooseker and DA Goodenough

A tight junction-enriched membrane fraction has been used as immunogen to generate a monoclonal antiserum specific for this intercellular junction. Hybridomas were screened for their ability to both react on an immunoblot and localize to the junctional complex region on frozen sections of unfixed mouse liver. A stable hybridoma line has been isolated that secretes an antibody (R26.4C) that localizes in thin section images of isolated mouse liver plasma membranes to the points of membrane contact at the tight junction. This antibody recognizes a polypeptide of approximately 225,000 D, detectable in whole liver homogenates as well as in the tight junction-enriched membrane fraction. R26.4C localizes to the junctional complex region of a number of other epithelia, including colon, kidney, and testis, and to arterial endothelium, as assayed by immunofluorescent staining of cryostat sections of whole tissue. This antibody also stains the junctional complex region in confluent monolayers of the Madin-Darby canine kidney epithelial cell line. Immunoblot analysis of Madin-Darby canine kidney cells demonstrates the presence of a polypeptide similar in molecular weight to that detected in liver, suggesting that this protein is potentially a ubiquitous component of all mammalian tight junctions. The 225-kD tight junction-associated polypeptide is termed "ZO-1."
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Noguchi, J., Toyama, Y., Yuasa, S., Kikuchi, K., Kaneko, H. (2002). Hereditary Defects in Both Germ Cells and the Blood-Testis Barrier System in as-Mutant Rats: Evidence from Spermatogonial Transplantation and Tracer-Permeability Analysis. Biol. Reprod. 67: 880-888 [Abstract] [Full Text]
Masuda, M., Yageta, M., Fukuhara, H., Kuramochi, M., Maruyama, T., Nomoto, A., Murakami, Y. (2002). The Tumor Suppressor Protein TSLC1 Is Involved in Cell-Cell Adhesion. J. Biol. Chem. 277: 31014-31019 [Abstract] [Full Text]
D'Atri, F., Nadalutti, F., Citi, S. (2002). Evidence for a Functional Interaction between Cingulin and ZO-1 in Cultured Cells. J. Biol. Chem. 277: 27757-27764 [Abstract] [Full Text]
Hirose, T., Izumi, Y., Nagashima, Y., Tamai-Nagai, Y., Kurihara, H., Sakai, T., Suzuki, Y., Yamanaka, T., Suzuki, A., Mizuno, K., Ohno, S. (2002). Involvement of ASIP/PAR-3 in the promotion of epithelial tight junction formation. J. Cell Sci. 115: 2485-2495 [Abstract] [Full Text]
Furuse, M., Hata, M., Furuse, K., Yoshida, Y., Haratake, A., Sugitani, Y., Noda, T., Kubo, A., Tsukita, S. (2002). Claudin-based tight junctions are crucial for the mammalian epidermal barrier: a lesson from claudin-1-deficient mice. JCB 156: 1099-1111 [Abstract] [Full Text]