Clustering of Nck by a 12-residue Tir phosphopeptide is sufficient to trigger localized actin assembly
Biochemistry & Molecular Pharmacology
Graduate School of Biomedical Sciences; Department of Molecular Genetics and Microbiology
Medical Subject Headings
Actinin; Actins; Adhesins, Bacterial; Animals; Cell Adhesion; Cell Membrane; Cytoskeleton; Escherichia coli; Escherichia coli Proteins; Hela Cells; Humans; Mice; Nerve Tissue Proteins; Oncogene Proteins; Oocytes; Phosphopeptides; Protein Conformation; Protein Structure, Tertiary; Receptors, Cell Surface; Recombinant Fusion Proteins; Signal Transduction; Wiskott-Aldrich Syndrome Protein, Neuronal; Xenopus
Life Sciences | Medicine and Health Sciences
Enteropathogenic Escherichia coli (EPEC) translocates effector proteins into mammalian cells to promote reorganization of the cytoskeleton into filamentous actin pedestals. One effector, Tir, is a transmembrane receptor for the bacterial surface adhesin intimin, and intimin binding by the extracellular domain of Tir is required for actin assembly. The cytoplasmic NH2 terminus of Tir interacts with focal adhesion proteins, and its tyrosine-phosphorylated COOH terminus binds Nck, a host adaptor protein critical for pedestal formation. To define the minimal requirements for EPEC-mediated actin assembly, Tir derivatives were expressed in mammalian cells in the absence of all other EPEC components. Replacement of the NH2 terminus of Tir with a viral membrane-targeting sequence promoted efficient surface expression of a COOH-terminal Tir fragment. Artificial clustering of this fusion protein revealed that the COOH terminus of Tir, by itself, is sufficient to initiate a complete signaling cascade leading to pedestal formation. Consistent with this finding, clustering of Nck by a 12-residue Tir phosphopeptide triggered actin tail formation in Xenopus egg extracts.
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Citation: J Cell Biol. 2004 Feb 2;164(3):407-16. Link to article on publisher's site
DOI of Published Version
The Journal of cell biology
Campellone, Kenneth Geno; Rankin, Susannah; Pawson, Tony; Kirschner, Marc W.; Tipper, Donald J.; and Leong, John M., "Clustering of Nck by a 12-residue Tir phosphopeptide is sufficient to trigger localized actin assembly" (2004). GSBS Student Publications. 177.