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Volume 151, Issue 9

Ca release from host intracellular stores and related signal transduction during 81-176 internalization into human intestinal cells Free

Abstract

is the leading bacterial cause of human diarrhoeal disease in many parts of the world, including the USA. The ability of to invade the host intestinal epithelium is an important determinant of virulence. A common theme among pathogenic invasive micro-organisms is their ability to usurp the eukaryotic cell-signalling systems both to allow for invasion and to trigger disease pathogenesis. Ca is very important in a great variety of eukaryotic cell-signalling processes (e.g. calmodulin-activated enzymes, nuclear transcriptional upregulation, and cytoskeletal rearrangements). This study analyses the effects of Ca availability on invasion of human INT407 intestinal epithelial cells by strain 81-176. The ability of to invade INT407 cells was not blocked by chelation of any remaining extracellular Ca from host cells incubated in Ca-free, serum-free media. In contrast, invasion was markedly reduced either by chelating host intracellular Ca with 1,2-bis-(2-)ethane-,,′,′-tetraacetic acid acetoxymethyl ester (BAPTA, AM) or by blocking the release of Ca from intracellular stores with dantrolene or U73122. Moreover, Bay K8644, a plasma-membrane Ca-channel agonist, was observed to stimulate invasion, presumably by increasing host intracellular free Ca levels. Measurement of host-cell cytosolic Ca via spectrofluorimetry and fluorescence microscopy revealed an increase in Ca from 10 min post-infection. Monolayer pretreatment with either a calmodulin antagonist or a specific inhibitor of protein kinase C was found to cause a marked reduction in invasion, suggesting roles for these Ca-activated modulators in signal-transduction events involved in invasion. These results demonstrate that induces the mobilization of Ca from host intracellular stores, which is an essential step in the invasion of intestinal cells by this pathogen.

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Keyword(s): AM, acetoxymethyl ester , BAPTA, 1,2-bis-(2-)ethane-N,N,N′,N′-tetraacetic acid , CaM, calmodulin , ER, endoplasmic reticulum , IP3, inositol triphosphate , PKC, protein kinase C and PLC, phospholipase C
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2005-09-01
2024-04-25
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Ca2+ release from host intracellular stores and related signal transduction during Campylobacter jejuni 81-176 internalization into human intestinal cells
Microbiology 151, 3097 (2005); https://doi.org/10.1099/mic.0.27866-0
/content/journal/micro/10.1099/mic.0.27866-0
/content/journal/micro/10.1099/mic.0.27866-0
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