Scientific article
Open access

Cingulin Contains Globular and Coiled-coil Domains and Interacts with ZO-1, ZO-2, ZO-3, and Myosin

Published inThe Journal of Cell Biology, vol. 147, no. 7, p. 1569-1581
Publication date1999

We characterized the sequence and protein interactions of cingulin, an Mr 140–160-kD phosphoprotein localized on the cytoplasmic surface of epithelial tight junctions (TJ). The derived amino acid sequence of a full-length Xenopus laevis cingulin cDNA shows globular head (residues 1–439) and tail (1,326–1,368) domains and a central α-helical rod domain (440–1,325). Sequence analysis, electron microscopy, and pull-down assays indicate that the cingulin rod is responsible for the formation of coiled-coil parallel dimers, which can further aggregate through intermolecular interactions. Pull-down assays from epithelial, insect cell, and reticulocyte lysates show that an NH2-terminal fragment of cingulin (1–378) interacts in vitro with ZO-1 (Kd ∼5 nM), ZO-2, ZO-3, myosin, and AF-6, but not with symplekin, and a COOH-terminal fragment (377–1,368) interacts with myosin and ZO-3. ZO-1 and ZO-2 immunoprecipitates contain cingulin, suggesting in vivo interactions. Full-length cingulin, but not NH2-terminal and COOH-terminal fragments, colocalizes with endogenous cingulin in transfected MDCK cells, indicating that sequences within both head and rod domains are required for TJ localization. We propose that cingulin is a functionally important component of TJ, linking the submembrane plaque domain of TJ to the actomyosin cytoskeleton.

  • Cingulin
  • Tight junction
  • Epithelia
  • MDCK
  • Protein
Citation (ISO format)
CORDENONSI, Michelangelo et al. Cingulin Contains Globular and Coiled-coil Domains and Interacts with ZO-1, ZO-2, ZO-3, and Myosin. In: The Journal of Cell Biology, 1999, vol. 147, n° 7, p. 1569–1581. doi: 10.1083/jcb.147.7.1569
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Article (Published version)
ISSN of the journal0021-9525

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