Protein kinase Cζ phosphorylates occludin and promotes assembly of epithelial tight junctions

Suneet Jain, Takuya Suzuki, Ankur Seth, Geetha Samak, Radhakrishna Rao

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Protein kinases play an important role in the regulation of epithelial tight junctions. In the present study, we investigated the role of PKCζ (protein kinase Cζ) in tight junction regulation in Caco-2 and MDCK (Madin-Darby canine kidney) cell monolayers. Inhibition of PKCζ by a specific PKCζ pseudosubstrate peptide results in redistribution of occludin and ZO-1 (zona occludens 1) from the intercellular junctions and disruption of barrier function without affecting cell viability. Reduced expression of PKCζ by antisense oligonucleotide or shRNA (short hairpin RNA) also results in compromised tight junction integrity. Inhibition or knockdown of PKCζ delays calcium-induced assembly of tight junctions. Tight junction disruption by PKCζ pseudosubstrate is associated with the dephosphorylation of occludin and ZO-1 on serine and threonine residues. PKCζ directly binds to the C-terminal domain of occludin and phosphorylates it on threonine residues. Thr 403, Thr 404, Thr 424 and Thr 438 in the occludin C-terminal domain are the predominant sites of PKCζ-dependent phosphorylation. A T424A or T438A mutation in full-length occludin delays its assembly into the tight junctions. Inhibition of PKCζ also induces redistribution of occludin and ZO-1 from the tight junctions and dissociates these proteins from the detergent-insoluble fractions in mouse ileum. The present study demonstrates that PKCζ phosphorylates occludin on specific threonine residues and promotes assembly of epithelial tight junctions.

Original languageEnglish (US)
Pages (from-to)289-299
Number of pages11
JournalBiochemical Journal
Volume437
Issue number2
DOIs
StatePublished - Jul 15 2011

Fingerprint

Occludin
Tight Junctions
Protein Kinase C
Herpes Zoster
Threonine
Zonula Occludens-1 Protein
Tight Junction Proteins
Phosphorylation
Madin Darby Canine Kidney Cells
Intercellular Junctions
Antisense Oligonucleotides
Ileum
Detergents
Protein Kinases
Serine
Small Interfering RNA
Monolayers
Cell Survival
Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Protein kinase Cζ phosphorylates occludin and promotes assembly of epithelial tight junctions. / Jain, Suneet; Suzuki, Takuya; Seth, Ankur; Samak, Geetha; Rao, Radhakrishna.

In: Biochemical Journal, Vol. 437, No. 2, 15.07.2011, p. 289-299.

Research output: Contribution to journalArticle

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