Distinct pathways of cell migration and antiapoptotic response to epithelial injury: Structure-function analysis of human intestinal trefoil factor

Koichi Kinoshita, Douglas R. Taupin, Hiroshi Itoh, Daniel K. Podolsky

Research output: Contribution to journalArticle

169 Citations (Scopus)

Abstract

The trefoil peptide intestinal trefoil factor (ITF) plays a critical role in the protection of colonic mucosa and is essential to restitution after epithelial damage. These functional properties are accomplished through coordinated promotion of cell migration and inhibition of apoptosis. ITF contains a unique three-looped trefoil motif formed by intrachain disulfide bonds among six conserved cysteine residues, which is thought to contribute to its marked protease resistance. ITF also has a seventh cysteine residue, which permits homodimer formation. A series of cysteine-to-serine substitutions and a C-terminally truncated ITF were made by PCR site-directed mutagenesis. Any alteration of the trefoil motif or truncation resulted in loss of protease resistance. However, neither an intact trefoil domain nor dimerization was required to promote cell migration. This pro-restitution activity correlated with the ability of the ITF mutants to activate mitogen- activated protein (MAP) kinase independent of phosphorylation of the epidermal growth factor (EGF) receptor. In contrast, only intact ITF retained both phosphatidylinositol 3-kinase and the EGF receptor-dependent antiapoptotic effect in HCT116 and IEC-6 cells. The inability to block apoptosis correlated with a loss of trefoil peptide-induced transactivation of the EGF receptor or Akt kinase in HT-29 cells. In addition to defining structural requirements for the functional properties of ITF, these findings demonstrate that distinct intracellular signaling pathways mediate the effects of ITF on cell migration and apoptosis.

Original languageEnglish (US)
Pages (from-to)4680-4690
Number of pages11
JournalMolecular and Cellular Biology
Volume20
Issue number13
DOIs
StatePublished - Jul 2000

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Cell Movement
Wounds and Injuries
Epidermal Growth Factor Receptor
Cysteine
Apoptosis
Peptide Hydrolases
Cell Migration Inhibition
Phosphatidylinositol 3-Kinase
HT29 Cells
Trefoil Factor-3
Dimerization
Site-Directed Mutagenesis
Mitogen-Activated Protein Kinases
Disulfides
Serine
Transcriptional Activation
Mucous Membrane
Phosphotransferases
Phosphorylation
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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Distinct pathways of cell migration and antiapoptotic response to epithelial injury : Structure-function analysis of human intestinal trefoil factor. / Kinoshita, Koichi; Taupin, Douglas R.; Itoh, Hiroshi; Podolsky, Daniel K.

In: Molecular and Cellular Biology, Vol. 20, No. 13, 07.2000, p. 4680-4690.

Research output: Contribution to journalArticle

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