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Volume 153, Issue 12

Invasion of HeLa cells by group B streptococcus requires the phosphoinositide-3-kinase signalling pathway and modulates phosphorylation of host-cell Akt and glycogen synthase kinase-3 Free

Abstract

The group B streptococcus (GBS) is an opportunistic bacterial pathogen with the ability to cause invasive disease. While the ability of GBS to invade a number of host-cell types has been clearly demonstrated, the invasion process is not well understood at the molecular level. What has been well established is that modulation of host-cell actin microfilaments is essential for GBS invasion to occur. Phosphoinositide-3 kinase (PI3K) is a key regulator of the cytoskeleton in eukaryotic cells. Our goal in this investigation was to explore the role of the PI3K/Akt signalling pathway in epithelial cell invasion by GBS. The epithelial cell invasion process was mimicked using the HeLa 229 cell-culture model. Treating HeLa cells with chemical inhibitors of PI3K, Akt or Ras prior to bacterial infection inhibited GBS invasion but not attachment; treatment with 30 μM LY294002 (PI3K inhibitor) reduced GBS invasion by 75 %, 20 μM -6-hydroxymethyl--inositol 2-(R)-2--methyl-3--octadecylcarbonate (ICIO) (Akt inhibitor) reduced GBS invasion by 50 %, and 10 μM manumycin A (Ras inhibitor) inhibited GBS invasion by 90 %. Genetic inactivation of the p85 or p110 PI3K subunits in HeLa cells also reduced GBS invasion by 55 and 30 %, respectively. Western blot analysis revealed that phosphorylation of host-cell Akt and glycogen synthase kinase-3 (GSK-3) occurs in response to GBS infection, and that this is mediated upstream by PI3K. Infection of HeLa cells with GBS triggers pro-survival signalling and protects the HeLa cells from camptothecin-induced caspase-3 cleavage. The results from this investigation show that GBS both requires and activates the PI3K/Akt host-cell signalling pathway during invasion of epithelial cells.

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Keyword(s): CPT, camptothecin , GAS, group A streptococcus/streptococci , GBS, group B streptococcus/streptococci , GSK-3, glycogen synthase kinase-3 , HK-GBS, heat-killed GBS , ICIO, l-6-hydroxymethyl-chiro-inositol 2-(R)-2-O-methyl-3-O-octadecylcarbonate , PDK1, phosphoinositide-dependent protein kinase 1 , PH, plesktrin homology , PI3K, phosphoinositide-3 kinase , PIP2, phosphatidylinositol 3,4-bisphosphate , PIP3, phosphatidylinositol 3,4,5-triphosphate and PKC, protein kinase C
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2007-12-01
2023-10-01
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Invasion of HeLa cells by group B streptococcus requires the phosphoinositide-3-kinase signalling pathway and modulates phosphorylation of host-cell Akt and glycogen synthase kinase-3
Microbiology 153, 4240 (2007); https://doi.org/10.1099/mic.0.2007/008417-0
/content/journal/micro/10.1099/mic.0.2007/008417-0
/content/journal/micro/10.1099/mic.0.2007/008417-0
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