Intracellular membrane localization of pseudomonas ExoS and Yersinia YopE in mammalian cells
| Autor: | Joseph T. Barbieri, Yue Zhang, Rebecca Krall |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Yersinia Infections
Bacterial Toxins Plasma protein binding CHO Cells Biology Biochemistry Protein structure Cricetinae Animals Humans Pseudomonas Infections Binding site Molecular Biology Sequence Deletion chemistry.chemical_classification ADP Ribose Transferases Binding Sites Chinese hamster ovary cell Cell Biology Intracellular Membranes Fusion protein Cell biology Amino acid Protein Structure Tertiary Membrane protein chemistry Intracellular Bacterial Outer Membrane Proteins HeLa Cells Protein Binding |
| Zdroj: | The Journal of biological chemistry. 279(4) |
| ISSN: | 0021-9258 |
| Popis: | ExoS (453 amino acids) is a bi-functional type-III cytotoxin of Pseudomonas aeruginosa. Residues 96-233 comprise the Rho GTPase-activating protein (Rho GAP) domain, while residues 234-453 comprise the 14-3-3-dependent ADP-ribosyltransferase domain. Residues 51-72 represent a membrane localization domain (MLD), which targets ExoS to perinuclear vesicles within mammalian cells. YopE (219 amino acids) is a type-III cytotoxin of Yersinia that is also a Rho GAP. Residues 96-219 comprise the YopE Rho GAP domain. While the Rho GAP domains of ExoS and YopE share structural homology, unlike ExoS, the intracellular localization of YopE within mammalian cells has not been resolved and is the subject of this investigation. Deletion mapping showed that the N terminus of YopE was required for intracellular membrane localization of YopE in CHO cells. A fusion protein containing the N-terminal 84 amino acids of YopE localized to a punctate-perinuclear region in mammalian cells and co-localized with a fusion protein containing the MLD of ExoS. Residues 54-75 of YopE (termed YopE-MLD) were necessary and sufficient for intracellular localization in mammalian cells. The YopE-MLD localized ExoS to intracellular membranes and targeted ExoS to ADP-ribosylate small molecular weight membrane proteins as observed for native type-III delivered ExoS. These data indicate that the YopE MLD functionally complements the ExoS MLD for intracellular targeting in mammalian cells. |
| Databáze: | OpenAIRE |
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