title: Differential phosphoproteome profiling reveals a functional role for VASP in Helicobacter pylori-induced cytoskeleton turnover in gastric epithelial cells.
abstract: 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.
authors.affiliation: Junior Research Group, Paul-Ehrlich Institute, Paul-Ehrlich Str. 51-59, D-63225 Langen, Germany.
authors.forename: Olivia
authors.initials: O
authors.lastname: Knauer
authors.forename: Nadine A
authors.initials: NA
authors.lastname: Binai
authors.forename: Gert
authors.initials: G
authors.lastname: Carra
authors.forename: Tobias
authors.initials: T
authors.lastname: Beckhaus
authors.forename: Kay-Martin
authors.initials: KM
authors.lastname: Hanschmann
authors.forename: Thomas
authors.initials: T
authors.lastname: Renné
authors.forename: Steffen
authors.initials: S
authors.lastname: Backert
authors.forename: Michael
authors.initials: M
authors.lastname: Karas
authors.forename: Silja
authors.initials: S
authors.lastname: Wessler
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism* / Q000382:microbiology; D005753:Gastric Mucosa / Q000166:cytology* / Q000378:metabolism / Q000382:microbiology; D016481:Helicobacter Infections / Q000378:metabolism*; D016480:Helicobacter pylori / Q000378:metabolism*; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C080866:cagA protein, Helicobacter pylori; D000098662:Vasodilator-Stimulated Phosphoprotein; D017869:Cyclic GMP-Dependent Protein Kinases