Contrasting effects of ERK on tight junction integrity in differentiated and under-differentiated Caco-2 cell monolayers

Sudhir Aggarwal, Takuya Suzuki, William L. Taylor, Aditi Bhargava, Radhakrishna Rao

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

ERK (extracellular-signal-regulated kinase) activation leads to disruption of tight junctions in some epithelial monolayers, whereas it prevents disruption of tight junctions in other epithelia. The factors responsible for such contrasting influences of ERK on tight junction integrity are unknown. The present study investigated the effect of the state of cell differentiation on ERK-mediated regulation of tight junctions in Caco-2 cell monolayers. EGF (epidermal growth factor) potentiated H2O2-induced tight junction disruption in under-differentiated cell monolayers, which was attenuated by the MEK [MAPK (mitogen-activated protein kinase)/ERK kinase] inhibitor U0126. In contrast, EGF prevented H2O2-induced disruption of tight junctions in differentiated cell monolayers, which was also attenuated by U0126. Knockdown of ERK1/2 enhanced tight junction integrity and accelerated assembly of tight junctions in under-differentiated cell monolayers, whereas it had the opposite effect in differentiated cell monolayers. Regulated expression of wild-type and constitutively activeMEK1 disrupted tight junctions, and the expression of dominant-negative MEK1 enhanced tight junction integrity in under-differentiated cells, whereas contrasting responses were recorded in differentiated cells. EGF prevented both H2O 2-induced association of PP2A (protein phosphatase 2A), and loss of association of PKCζ (protein kinase Cζ), with occludin by an ERK-dependent mechanism in differentiated cell monolayers, but not in underdifferentiated cell monolayers. Active ERK was distributed in the intracellular compartment in under-differentiated cell monolayers, whereas it was localized mainly in the perijunctional region in differentiated cell monolayers. Thus ERK may exhibit its contrasting influences on tight junction integrity in underdifferentiated and differentiated epithelial cells by virtue of differences in its subcellular distribution and ability to regulate the association of PKCζ; and PP2A with tight junction proteins.

Original languageEnglish (US)
Pages (from-to)51-63
Number of pages13
JournalBiochemical Journal
Volume433
Issue number1
DOIs
StatePublished - Jan 1 2011

Fingerprint

Caco-2 Cells
Tight Junctions
Extracellular Signal-Regulated MAP Kinases
Monolayers
Epidermal Growth Factor
Protein Phosphatase 2
Protein Kinase C
Association reactions
Occludin
Tight Junction Proteins
Mitogen-Activated Protein Kinase Kinases
Mitogen-Activated Protein Kinases
Phosphotransferases
Cell Differentiation
Chemical activation
Epithelium
Epithelial Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Contrasting effects of ERK on tight junction integrity in differentiated and under-differentiated Caco-2 cell monolayers. / Aggarwal, Sudhir; Suzuki, Takuya; Taylor, William L.; Bhargava, Aditi; Rao, Radhakrishna.

In: Biochemical Journal, Vol. 433, No. 1, 01.01.2011, p. 51-63.

Research output: Contribution to journalArticle

Aggarwal, Sudhir ; Suzuki, Takuya ; Taylor, William L. ; Bhargava, Aditi ; Rao, Radhakrishna. / Contrasting effects of ERK on tight junction integrity in differentiated and under-differentiated Caco-2 cell monolayers. In: Biochemical Journal. 2011 ; Vol. 433, No. 1. pp. 51-63.
@article{4f192b58f0c94329b92964bd8e0a720c,
title = "Contrasting effects of ERK on tight junction integrity in differentiated and under-differentiated Caco-2 cell monolayers",
abstract = "ERK (extracellular-signal-regulated kinase) activation leads to disruption of tight junctions in some epithelial monolayers, whereas it prevents disruption of tight junctions in other epithelia. The factors responsible for such contrasting influences of ERK on tight junction integrity are unknown. The present study investigated the effect of the state of cell differentiation on ERK-mediated regulation of tight junctions in Caco-2 cell monolayers. EGF (epidermal growth factor) potentiated H2O2-induced tight junction disruption in under-differentiated cell monolayers, which was attenuated by the MEK [MAPK (mitogen-activated protein kinase)/ERK kinase] inhibitor U0126. In contrast, EGF prevented H2O2-induced disruption of tight junctions in differentiated cell monolayers, which was also attenuated by U0126. Knockdown of ERK1/2 enhanced tight junction integrity and accelerated assembly of tight junctions in under-differentiated cell monolayers, whereas it had the opposite effect in differentiated cell monolayers. Regulated expression of wild-type and constitutively activeMEK1 disrupted tight junctions, and the expression of dominant-negative MEK1 enhanced tight junction integrity in under-differentiated cells, whereas contrasting responses were recorded in differentiated cells. EGF prevented both H2O 2-induced association of PP2A (protein phosphatase 2A), and loss of association of PKCζ (protein kinase Cζ), with occludin by an ERK-dependent mechanism in differentiated cell monolayers, but not in underdifferentiated cell monolayers. Active ERK was distributed in the intracellular compartment in under-differentiated cell monolayers, whereas it was localized mainly in the perijunctional region in differentiated cell monolayers. Thus ERK may exhibit its contrasting influences on tight junction integrity in underdifferentiated and differentiated epithelial cells by virtue of differences in its subcellular distribution and ability to regulate the association of PKCζ; and PP2A with tight junction proteins.",
author = "Sudhir Aggarwal and Takuya Suzuki and Taylor, {William L.} and Aditi Bhargava and Radhakrishna Rao",
year = "2011",
month = "1",
day = "1",
doi = "10.1042/BJ20100249",
language = "English (US)",
volume = "433",
pages = "51--63",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "1",

}

TY - JOUR

T1 - Contrasting effects of ERK on tight junction integrity in differentiated and under-differentiated Caco-2 cell monolayers

AU - Aggarwal, Sudhir

AU - Suzuki, Takuya

AU - Taylor, William L.

AU - Bhargava, Aditi

AU - Rao, Radhakrishna

PY - 2011/1/1

Y1 - 2011/1/1

N2 - ERK (extracellular-signal-regulated kinase) activation leads to disruption of tight junctions in some epithelial monolayers, whereas it prevents disruption of tight junctions in other epithelia. The factors responsible for such contrasting influences of ERK on tight junction integrity are unknown. The present study investigated the effect of the state of cell differentiation on ERK-mediated regulation of tight junctions in Caco-2 cell monolayers. EGF (epidermal growth factor) potentiated H2O2-induced tight junction disruption in under-differentiated cell monolayers, which was attenuated by the MEK [MAPK (mitogen-activated protein kinase)/ERK kinase] inhibitor U0126. In contrast, EGF prevented H2O2-induced disruption of tight junctions in differentiated cell monolayers, which was also attenuated by U0126. Knockdown of ERK1/2 enhanced tight junction integrity and accelerated assembly of tight junctions in under-differentiated cell monolayers, whereas it had the opposite effect in differentiated cell monolayers. Regulated expression of wild-type and constitutively activeMEK1 disrupted tight junctions, and the expression of dominant-negative MEK1 enhanced tight junction integrity in under-differentiated cells, whereas contrasting responses were recorded in differentiated cells. EGF prevented both H2O 2-induced association of PP2A (protein phosphatase 2A), and loss of association of PKCζ (protein kinase Cζ), with occludin by an ERK-dependent mechanism in differentiated cell monolayers, but not in underdifferentiated cell monolayers. Active ERK was distributed in the intracellular compartment in under-differentiated cell monolayers, whereas it was localized mainly in the perijunctional region in differentiated cell monolayers. Thus ERK may exhibit its contrasting influences on tight junction integrity in underdifferentiated and differentiated epithelial cells by virtue of differences in its subcellular distribution and ability to regulate the association of PKCζ; and PP2A with tight junction proteins.

AB - ERK (extracellular-signal-regulated kinase) activation leads to disruption of tight junctions in some epithelial monolayers, whereas it prevents disruption of tight junctions in other epithelia. The factors responsible for such contrasting influences of ERK on tight junction integrity are unknown. The present study investigated the effect of the state of cell differentiation on ERK-mediated regulation of tight junctions in Caco-2 cell monolayers. EGF (epidermal growth factor) potentiated H2O2-induced tight junction disruption in under-differentiated cell monolayers, which was attenuated by the MEK [MAPK (mitogen-activated protein kinase)/ERK kinase] inhibitor U0126. In contrast, EGF prevented H2O2-induced disruption of tight junctions in differentiated cell monolayers, which was also attenuated by U0126. Knockdown of ERK1/2 enhanced tight junction integrity and accelerated assembly of tight junctions in under-differentiated cell monolayers, whereas it had the opposite effect in differentiated cell monolayers. Regulated expression of wild-type and constitutively activeMEK1 disrupted tight junctions, and the expression of dominant-negative MEK1 enhanced tight junction integrity in under-differentiated cells, whereas contrasting responses were recorded in differentiated cells. EGF prevented both H2O 2-induced association of PP2A (protein phosphatase 2A), and loss of association of PKCζ (protein kinase Cζ), with occludin by an ERK-dependent mechanism in differentiated cell monolayers, but not in underdifferentiated cell monolayers. Active ERK was distributed in the intracellular compartment in under-differentiated cell monolayers, whereas it was localized mainly in the perijunctional region in differentiated cell monolayers. Thus ERK may exhibit its contrasting influences on tight junction integrity in underdifferentiated and differentiated epithelial cells by virtue of differences in its subcellular distribution and ability to regulate the association of PKCζ; and PP2A with tight junction proteins.

UR - http://www.scopus.com/inward/record.url?scp=79551482121&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79551482121&partnerID=8YFLogxK

U2 - 10.1042/BJ20100249

DO - 10.1042/BJ20100249

M3 - Article

VL - 433

SP - 51

EP - 63

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 1

ER -