R J Haines1,2, R S Beard3, L Chen1, R A Eitnier1, M H Wu4,5. 1. Department of Surgery, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., MDC 8, Office MDC 2012, Tampa, FL, 33612, USA. 2. James A. Haley Veterans' Hospital, Tampa, FL, USA. 3. Department of Molecular Pharmacology and Physiology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. 4. Department of Surgery, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., MDC 8, Office MDC 2012, Tampa, FL, 33612, USA. mwu1@health.usf.edu. 5. James A. Haley Veterans' Hospital, Tampa, FL, USA. mwu1@health.usf.edu.
Abstract
BACKGROUND: IL-1β is a cytokine involved in mediating epithelial barrier dysfunction in the gut. It is known that IL-1β mediates activation of non-muscle myosin light chain kinase in epithelial cells, but the precise mechanism by which epithelial barrier dysfunction is induced by IL-1β is not understood. METHODS AND RESULTS: Using a Caco2 cell model, we show that the expression of the tight junction protein, claudin-3, is transcriptionally downregulated by IL-1β treatment. In addition, after assessing protein and mRNA expression, and protein localization, we show that inhibition of nmMLCK rescues IL-1β-mediated decrease in claudin-3 expression as well as junction protein redistribution. Using chromatin immunoprecipitation assays, we also show that β-catenin targeting of the claudin-3 promoter occurs as a consequence of IL-1β-mediated epithelial barrier dysfunction, and inhibition of nmMLCK interferes with this interaction. CONCLUSIONS: Taken together, these data represent the first line of evidence demonstrating nmMLCK regulation of claudin-3 expression in response to IL-1β-treated epithelial cells.
BACKGROUND: IL-1β is a cytokine involved in mediating epithelial barrier dysfunction in the gut. It is known that IL-1β mediates activation of non-muscle myosin light chain kinase in epithelial cells, but the precise mechanism by which epithelial barrier dysfunction is induced by IL-1β is not understood. METHODS AND RESULTS: Using a Caco2 cell model, we show that the expression of the tight junction protein, claudin-3, is transcriptionally downregulated by IL-1β treatment. In addition, after assessing protein and mRNA expression, and protein localization, we show that inhibition of nmMLCK rescues IL-1β-mediated decrease in claudin-3 expression as well as junction protein redistribution. Using chromatin immunoprecipitation assays, we also show that β-catenin targeting of the claudin-3 promoter occurs as a consequence of IL-1β-mediated epithelial barrier dysfunction, and inhibition of nmMLCK interferes with this interaction. CONCLUSIONS: Taken together, these data represent the first line of evidence demonstrating nmMLCK regulation of claudin-3 expression in response to IL-1β-treated epithelial cells.
Authors: Shyam Prasad; Roberto Mingrino; Katri Kaukinen; Katherine L Hayes; Robert M Powell; Thomas T MacDonald; Jane E Collins Journal: Lab Invest Date: 2005-09 Impact factor: 5.662
Authors: S Zeissig; N Bürgel; D Günzel; J Richter; J Mankertz; U Wahnschaffe; A J Kroesen; M Zeitz; M Fromm; J-D Schulzke Journal: Gut Date: 2006-07-05 Impact factor: 23.059
Authors: Daniel R Clayburgh; Shari Rosen; Edwina D Witkowski; Fengjun Wang; Stephanie Blair; Steven Dudek; Joe G N Garcia; John C Alverdy; Jerrold R Turner Journal: J Biol Chem Date: 2004-10-26 Impact factor: 5.157
Authors: R J Haines; C Y Wang; C G Y Yang; R A Eitnier; F Wang; M H Wu Journal: Am J Physiol Gastrointest Liver Physiol Date: 2017-08-24 Impact factor: 4.052