PMID:11994151
| Citation |
Walburger, A, Lazdunski, C and Corda, Y (2002) The Tol/Pal system function requires an interaction between the C-terminal domain of TolA and the N-terminal domain of TolB. Mol. Microbiol. 44:695-708 |
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| Abstract |
The Tol/Pal system of Escherichia coli is composed of the YbgC, TolQ, TolA, TolR, TolB, Pal and YbgF proteins. It is involved in maintaining the integrity of the outer membrane, and is required for the uptake of group A colicins and DNA of filamentous bacteriophages. To identify new interactions between the components of the Tol/Pal system and gain insight into the mechanism of colicin import, we performed a yeast two-hybrid screen using the different components of the Tol/Pal system and colicin A. Using this system, we confirmed the already known interactions and identified several new interactions. TolB dimerizes and the periplasmic domain of TolA interacts with YbgF and TolB. Our results indicate that the central domain of TolA (TolAII) is sufficient to interact with YbgF, that the C-terminal domain of TolA (TolAIII) is sufficient to interact with TolB, and that the amino terminal domain of TolB (D1) is sufficient to bind TolAIII. The TolA/TolB interaction was confirmed by cross-linking experiments on purified proteins. Moreover, we show that the interaction between TolA and TolB is required for the uptake of colicin A and for the membrane integrity. These results demonstrate that the TolA/TolB interaction allows the formation of a trans-envelope complex that brings the inner and outer membranes in close proximity. |
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| Keywords |
Bacterial Outer Membrane Proteins/chemistry; Bacterial Outer Membrane Proteins/physiology; Bacterial Proteins/genetics; Biological Transport; Cell Membrane/chemistry; Cell Membrane/drug effects; Cell Membrane/ultrastructure; Colicins/metabolism; Cross-Linking Reagents/pharmacology; Escherichia coli/drug effects; Escherichia coli/physiology; Escherichia coli/ultrastructure; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/pharmacology; Escherichia coli Proteins/physiology; Formaldehyde/pharmacology; Lipoproteins/chemistry; Lipoproteins/physiology; Macromolecular Substances; Membrane Proteins/chemistry; Membrane Proteins/pharmacology; Membrane Proteins/physiology; Mutagenesis; Peptidoglycan/chemistry; Polymerase Chain Reaction; Protein Interaction Mapping; Protein Structure, Tertiary; Proteoglycans; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/metabolism; Serine Endopeptidases/genetics; Sodium Dodecyl Sulfate/pharmacology; Two-Hybrid System Techniques |
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