Differential phosphoproteome profiling reveals a functional role for VASP in Helicobacter pylori-induced cytoskeleton turnover in gastric epithelial cells

TY - JOUR

T1 - Differential phosphoproteome profiling reveals a functional role for VASP in Helicobacter pylori-induced cytoskeleton turnover in gastric epithelial cells

AU - Knauer, Olivia

AU - Binai, Nadine A

AU - Carra, Gert

AU - Beckhaus, Tobias

AU - Hanschmann, Kay-Martin

AU - Renné, Thomas

AU - Backert, Steffen

AU - Karas, Michael

AU - Wessler, Silja

PY - 2008/11/1

Y1 - 2008/11/1

N2 - Infection with Helicobacter pylori induces various gastric diseases, including ulceration, gastritis and neoplasia. As H. pylori-induced cellular mechanisms leading to these disease states are widely unclear, we analysed the phosphoproteome of H. pylori-infected gastric epithelial cells. Phosphoproteins from infected cells were enriched using affinity columns and analysed by two-dimensional gel electrophoresis and mass spectrometry. Eleven novel phosphoproteins that showed differentially regulated phosphorylation levels during H. pylori infection were identified. Interestingly, the identified proteins were actin-binding, transport and folding, RNA/DNA-binding or cancer-associated proteins. We analysed functions of one identified H. pylori-regulated candidate, the vasodilator-stimulated phosphoprotein (VASP). H. pylori induced VASP phosphorylation at residues Ser157, Ser239 and Thr278, which was enhanced by the bacterial oncogene cytotoxin-associated gene A. Overexpression of a phosphorylation-resistant VASP mutant efficiently blocked host cell elongation. We identified cGMP-dependent protein kinase G-mediated Ser239 and Thr278 phosphorylation of VASP as a crucial event in H. pylori-dependent host cell elongation. These results suggest that phosphorylated VASP could be a novel target candidate for therapeutic intervention in H. pylori-related gastric diseases.

AB - Infection with Helicobacter pylori induces various gastric diseases, including ulceration, gastritis and neoplasia. As H. pylori-induced cellular mechanisms leading to these disease states are widely unclear, we analysed the phosphoproteome of H. pylori-infected gastric epithelial cells. Phosphoproteins from infected cells were enriched using affinity columns and analysed by two-dimensional gel electrophoresis and mass spectrometry. Eleven novel phosphoproteins that showed differentially regulated phosphorylation levels during H. pylori infection were identified. Interestingly, the identified proteins were actin-binding, transport and folding, RNA/DNA-binding or cancer-associated proteins. We analysed functions of one identified H. pylori-regulated candidate, the vasodilator-stimulated phosphoprotein (VASP). H. pylori induced VASP phosphorylation at residues Ser157, Ser239 and Thr278, which was enhanced by the bacterial oncogene cytotoxin-associated gene A. Overexpression of a phosphorylation-resistant VASP mutant efficiently blocked host cell elongation. We identified cGMP-dependent protein kinase G-mediated Ser239 and Thr278 phosphorylation of VASP as a crucial event in H. pylori-dependent host cell elongation. These results suggest that phosphorylated VASP could be a novel target candidate for therapeutic intervention in H. pylori-related gastric diseases.

KW - Animals

KW - Antigens, Bacterial

KW - Bacterial Proteins

KW - Cell Adhesion Molecules

KW - Cell Line

KW - Cyclic GMP-Dependent Protein Kinases

KW - Cytoskeleton

KW - Epithelial Cells

KW - Gastric Mucosa

KW - Helicobacter Infections

KW - Helicobacter pylori

KW - Humans

KW - Microfilament Proteins

KW - Phosphoproteins

KW - Recombinant Fusion Proteins

KW - Signal Transduction

U2 - 10.1111/j.1462-5822.2008.01207.x

DO - 10.1111/j.1462-5822.2008.01207.x

M3 - SCORING: Journal article

C2 - 18637808

VL - 10

SP - 2285

EP - 2296

JO - CELL MICROBIOL

JF - CELL MICROBIOL

SN - 1462-5814

IS - 11

ER -