title: EPIYA motif is a membrane-targeting signal of Helicobacter pylori virulence factor CagA in mammalian cells.
abstract: Helicobacter pylori contributes to the development of peptic ulcers and atrophic gastritis. Furthermore, H. pylori strains carrying the cagA gene are more virulent than cagA-negative strains and are associated with the development of gastric adenocarcinoma. The cagA gene product, CagA, is translocated into gastric epithelial cells and localizes to the inner surface of the plasma membrane, in which it undergoes tyrosine phosphorylation at the Glu-Pro-Ile-Tyr-Ala (EPIYA) motif. Tyrosine-phosphorylated CagA specifically binds to and activates Src homology 2-containing protein-tyrosine phosphatase-2 (SHP-2) at the membrane, thereby inducing an elongated cell shape termed the hummingbird phenotype. Accordingly, membrane tethering of CagA is an essential prerequisite for the pathogenic activity of CagA. We show here that membrane association of CagA requires the EPIYA-containing region but is independent of EPIYA tyrosine phosphorylation. We further show that specific deletion of the EPIYA motif abolishes the ability of CagA to associate with the membrane. Conversely, reintroduction of an EPIYA sequence into a CagA mutant that lacks the EPIYA-containing region restores membrane association of CagA. Thus, the presence of a single EPIYA motif is necessary for the membrane localization of CagA. Our results indicate that the EPIYA motif has a dual function in membrane association and tyrosine phosphorylation, both of which are critically involved in the activity of CagA to deregulate intracellular signaling, and suggest that the EPIYA motif is a crucial therapeutic target of cagA-positive H. pylori infection.
authors.affiliation: Division of Molecular Oncology, Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Higashi
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Yokoyama
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Fujii
authors.forename: Shumei
authors.initials: S
authors.lastname: Ren
authors.forename: Hitomi
authors.initials: H
authors.lastname: Yuasa
authors.forename: Iraj
authors.initials: I
authors.lastname: Saadat
authors.forename: Naoko
authors.initials: N
authors.lastname: Murata-Kamiya
authors.forename: Takeshi
authors.initials: T
authors.lastname: Azuma
authors.forename: Masanori
authors.initials: M
authors.lastname: Hatakeyama
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000737:chemistry* / Q000502:physiology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000502:physiology*; D001665:Binding Sites; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000382:microbiology; D003593:Cytoplasm / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D005753:Gastric Mucosa / Q000378:metabolism; D005822:Genetic Vectors; D016480:Helicobacter pylori / Q000378:metabolism* / Q000472:pathogenicity*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010766:Phosphorylation; D011485:Protein Binding; D054648:Protein Phosphatase 2; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry; D011995:Recombination, Genetic; D015398:Signal Transduction; D014162:Transfection; D014443:Tyrosine / Q000737:chemistry; D014774:Virulence
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D047908:Intracellular Signaling Peptides and Proteins; C080866:cagA protein, Helicobacter pylori; D014443:Tyrosine; D054648:Protein Phosphatase 2; C516724:PTPN11 protein, human; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D017027:Protein Tyrosine Phosphatases