RRC ID 65745
Author Sougleri IS, Papadakos KS, Zadik MP, Mavri-Vavagianni M, Mentis AF, Sgouras DN.
Title Helicobacter pylori CagA protein induces factors involved in the epithelial to mesenchymal transition (EMT) in infected gastric epithelial cells in an EPIYA- phosphorylation-dependent manner.
Journal FEBS J
Abstract As a result of Helicobacter pylori adhesion to gastric epithelial cells, the bacterial effector cytotoxin-associated gene A (CagA) is translocated intracellularly, and after hierarchical tyrosine phosphorylation on multiple EPIYA motifs, de-regulates cellular polarity and contributes to induction of an elongation and scattering phenotype that resembles the epithelial to mesenchymal transition (EMT). Stromelysin-1/matrix metalloproteinase-3 (MMP-3) has been reported to induce a sequence of molecular alterations leading to stable EMT transition and carcinogenesis in epithelial cells. To identify the putative role of CagA protein in MMP-3 induction, we exploited an experimental H. pylori infection system in gastric epithelial cell lines. We utilized isogenic mutants expressing CagA protein with variable numbers of EPIYA and phosphorylation-deficient EPIFA motifs, as well as cagA knockout and translocation-deficient cagE knockout strains. Increased levels of MMP-3 transcriptional activation were demonstrated by quantitative real time-PCR for strains with more than two terminal EPIYA phosphorylation motifs in CagA. MMP-3 expression in total cell lysates and the corresponding culture supernatants was associated with CagA expression and translocation and was dependent on CagA phosphorylation. A CagA EPIYA phosphorylation-dependent increase in gelatinase and caseinolytic activity was also detected in culture supernatants by zymography. A significant increase in the transcriptional activity of the mesenchymal markers Vimentin, Snail and ZEB1 and the stem cell marker CD44 was observed in the case of CagA containing phosphorylation-functional EPIYA motifs. Our data suggest that CagA protein induces EMT through EPIYA phosphorylation-dependent up-regulation of MMP-3. Moreover, no significant increase in EMT and stem cell markers was observed following infection with H. pylori strains that cannot effectively translocate CagA protein.
Volume 283(2)
Pages 206-20
Published 2016-1-1
DOI 10.1111/febs.13592
PMID 26907789
MeSH Amino Acid Motifs Amino Acid Sequence Animals Antigens, Bacterial / genetics Antigens, Bacterial / metabolism* Bacterial Proteins / genetics Bacterial Proteins / metabolism* Cell Line, Tumor Epithelial Cells / metabolism* Epithelial Cells / microbiology Epithelial-Mesenchymal Transition* Gastric Mucosa / metabolism* Gastric Mucosa / microbiology Helicobacter Infections / metabolism* Helicobacter Infections / pathology Helicobacter pylori / pathogenicity* Host-Pathogen Interactions Humans Hyaluronan Receptors / metabolism Matrix Metalloproteinase 3 / metabolism Molecular Sequence Data Phosphorylation Vimentin / metabolism
IF 4.392
Human and Animal Cells MKN45(RCB1001)