Mechanism of extracellular calcium regulation of intestinal epithelial tight junction permeability: Role of cytoskeletal involvement

Thomas, Y.; Dat, Tran Do; Hoa, Neil; Nguyen, Don; Merryfield, Margaret; Tarnawski, Andrzej S.

doi:10.1002/1097-0029(20001015)51:2<156::aid-jemt7>3.0.co;2-j

Cited by 126 publications

(89 citation statements)

References 38 publications

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“…How certain second messengers such as intracellular calcium levels induce tight junction dysfunction is currently the subject of much research (61,62). In fact, intracellular calcium changes regulate the function of occludin through regulating occludin dephosphorylation and ubiquitination (63). ALO-induced increase of intracellular calcium to alter tight junction architecture could benefit movement of the vegetative anthrax bacteria or other bacterial toxins through this barrier for systemic infection.…”

Section: Discussionmentioning

confidence: 99%

Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis

Bourdeau

Malito

Chenal

et al. 2009

Journal of Biological Chemistry

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Anthrolysin O (ALO) is a pore-forming, cholesterol-dependent cytolysin (CDC) secreted by Bacillus anthracis, the etiologic agent for anthrax. Growing evidence suggests the involvement of ALO in anthrax pathogenesis. Here, we show that the apical application of ALO decreases the barrier function of human polarized epithelial cells as well as increases intracellular calcium and the internalization of the tight junction protein occludin. Using pharmacological agents, we also found that barrier function disruption requires increased intracellular calcium and protein degradation. We also report a crystal structure of the soluble state of ALO. Based on our analytical ultracentrifugation and light scattering studies, ALO exists as a monomer. Our ALO structure provides the molecular basis as to how ALO is locked in a monomeric state, in contrast to other CDCs that undergo antiparallel dimerization or higher order oligomerization in solution. ALO has four domains and is globally similar to perfringolysin O (PFO) and intermedilysin (ILY), yet the highly conserved undecapeptide region in domain 4 (D4) adopts a completely different conformation in all three CDCs. Consistent with the differences within D4 and at the D2-D4 interface, we found that ALO D4 plays a key role in affecting the barrier function of C2BBE cells, whereas PFO domain 4 cannot substitute for this role. Novel structural elements and unique cellular functions of ALO revealed by our studies provide new insight into the molecular basis for the diverse nature of the CDC family.Cholesterol-dependent cytolysins (CDCs) 4 are a family of pore-forming toxins from many organisms, including but not limited to the genera Archanobacterium, Bacillus, Clostridium, Listeria, and Streptococcus. Recently, work in vertebrates has revealed that CDCs and membrane attack complex/perforin superfamily domain-containing proteins share a similar fold, suggesting that vertebrates use a similar mechanism for defense against infection (1, 2). A common feature of the CDC family is the requirement of cholesterol in the membrane to form pores (3). In addition to cholesterol, certain members of the family also require a cellular receptor, such as CD59 for the toxin ILY from Streptococcus intermedius (4). The specific mechanism by which CDCs form pores is not completely resolved; however, what is generally known is that ring-shaped oligomerization at the cellular membrane is followed by large conformational changes in each unit of the oligomer, resulting in the insertion of a ␤-barrel into the cellular membrane (5). Pore formation results in a variety of downstream signaling effects, including but not limited to the influx of Ca 2ϩ into the cell (6).A good deal is known about structures of the prepore conformation of CDCs. The crystal structures of prepore PFO, from Clostridium perfringens, and ILY have previously been elucidated (7,8). Each structure shows a characteristic fourdomain architecture, in which domain 4 (D4) is involved in membrane recognition, domain 3 (D3) is involved in ␤...

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Section: Discussionmentioning

confidence: 99%

Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis

Bourdeau

Malito

Chenal

et al. 2009

Journal of Biological Chemistry

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“…The actin-myosin filaments of the pig epiblast cell's adhesion belts may place continuous contractile tension on the cell's apical portion (Hagmann et al, 1999;Langanger et al, 1986;Odell et al, 1981). Purse string-like contraction of the epiblast cell adhesion belts is also a possibility in response to the removal of external calcium because this has been shown to occur in Madin-Darby canine kidney (MDCK) epithelial monolayers (Castillo et al, 1998;Lagunes et al, 1999;Ma et al, 2000). Such tension and contraction could exacerbate cell rupture at the epiblast cell's basolateral desmosomes as external Ca ϩϩ is withdrawn and Ca ϩϩ -dependent cadherins, i.e., desmocollins and desmoglein, denature and dissociate (Collins and Fleming, 1995).…”

Section: Discussionmentioning

confidence: 99%

Ultrastructure of the embryonic stem cells of the 8‐day pig blastocyst before and after in vitro manipulation: Development of junctional apparatus and the lethal effects of PBS mediated cell–cell dissociation

Talbot

Garrett

2001

Anat. Rec.

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Ultrastructural examination of 8-day hatched pig blastocysts (large and small), their cultured inner cell mass (ICM), and cultured epiblast tissue (embryonic stem cells) was undertaken to assess the development of epiblast cell junctions and cytoskeletal elements. In small blastocysts, epiblast cells had no desmosomes or tight junction (TJ) connections and few organized microfilament bundles, whereas in large blastocysts the epiblast cells were connected by TJ and desmosomes with associated microfilaments. ICM isolation by immunodissection damaged the endoderm cells beneath the trophectoderm cells but did not appear to damage the epiblast cells or their associated endoderm cells. Epiblast cells in cultured ICMs were similar in character to those in the intact large blastocyst except that perinuclear microfilaments were observed. Isolated pig epiblasts, cultured for ϳ36 hr on STO feeder layers, formed a monolayer whose cells were connected by TJ, adherens junctions and desmosomes with prominent microfilament bundles running parallel to the apical cytoplasmic membranes. Perinuclear microfilaments were a consistent feature in the ϳ36 hr cultured epiblast cells. A feature characteristic of differentiation into notochordal cells, i.e., a solitary cilium, was also observed in the cultured epiblast. Exposure of the cultured epiblast cells to Ca ϩϩ -Mg ϩϩ -free phosphate buffered saline (PBS) for 5-10 min resulted in extensive cell blebbing and lysis. The results may indicate that pig epiblast cells could be more easily dissociated from early blastocysts (ϳ400 m in diameter) if immunodissection damage to the ICM can be avoided. It may be difficult, however, to establish them as embryonic stem cell lines because the cultured pig epiblast cells were easily lysed by standard cell-cell dissociation methods.

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“…Previous studies (17,(35)(36)(37)(38)(39) have shown that myosin L chain kinase (MLCK) plays a central role in the regulation of intestinal TJ permeability. Studies from our laboratory and others have shown that MLCK mediates pharmacologic (cytochalasins, ethanol, and low extracellular Ca ϩϩ solution) (37, 39 -41), physiologic (Na ϩ -glucose co-transport) (42), and microbial pathogen (enteropathogenic E. coli and C. difficile) (15, 43, 44) induced increase in intestinal TJ permeability.…”

Section: T He Defective Intestinal Epithelial Tight Junction (Tj)mentioning

confidence: 99%

Mechanism of IL-1β-Induced Increase in Intestinal Epithelial Tight Junction Permeability

Al–Sadi

Dokładny

et al. 2008

The Journal of Immunology

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361

295

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The IL-1β-induced increase in intestinal epithelial tight junction (TJ) permeability has been postulated to be an important mechanism contributing to intestinal inflammation of Crohn’s disease and other inflammatory conditions of the gut. The intracellular and molecular mechanisms that mediate the IL-1β-induced increase in intestinal TJ permeability remain unclear. The purpose of this study was to elucidate the mechanisms that mediate the IL-1β-induced increase in intestinal TJ permeability. Specifically, the role of myosin L chain kinase (MLCK) was investigated. IL-1β caused a progressive increase in MLCK protein expression. The time course of IL-1β-induced increase in MLCK level correlated linearly with increase in Caco-2 TJ permeability. Inhibition of the IL-1β-induced increase in MLCK protein expression prevented the increase in Caco-2 TJ permeability. Inhibition of the IL-1β-induced increase in MLCK activity also prevented the increase in Caco-2 TJ permeability. Additionally, knock-down of MLCK protein expression by small interference RNA prevented the IL-1β-induced increase in Caco-2 TJ permeability. The IL-1β-induced increase in MLCK protein expression was preceded by an increase in MLCK mRNA expression. The IL-1β-induced increase in MLCK mRNA transcription and subsequent increase in MLCK protein expression and Caco-2 TJ permeability was mediated by activation of NF-κB. In conclusion, our data indicate that the IL-1β increase in Caco-2 TJ permeability was mediated by an increase in MLCK expression and activity. Our findings also indicate that the IL-1β-induced increase in MLCK protein expression and Caco-2 TJ permeability was mediated by an NF-κB-dependent increase in MLCK gene transcription.

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Mechanism of extracellular calcium regulation of intestinal epithelial tight junction permeability: Role of cytoskeletal involvement

Cited by 126 publications

References 38 publications

Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis

Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis

Ultrastructure of the embryonic stem cells of the 8‐day pig blastocyst before and after in vitro manipulation: Development of junctional apparatus and the lethal effects of PBS mediated cell–cell dissociation

Mechanism of IL-1β-Induced Increase in Intestinal Epithelial Tight Junction Permeability

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