PMID:9620961
| Citation |
Cosma, CL, Crotwell, MD, Burrows, SY and Silhavy, TJ (1998) Folding-based suppression of extracytoplasmic toxicity conferred by processing-defective LamB. J. Bacteriol. 180:3120-30 |
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| Abstract |
We have utilized processing-defective derivatives of the outer membrane maltoporin, LamB, to study protein trafficking functions in the cell envelope of Escherichia coli. Our model proteins contain amino acid substitutions in the consensus site for cleavage by signal peptidase. As a result, the signal sequence is cleaved with reduced efficiency, effectively tethering the precursor protein to the inner membrane. These mutant porins are toxic when secreted to the cell envelope. Furthermore, strains producing these proteins exhibit altered outer membrane permeability, suggesting that the toxicity stems from some perturbation of the cell envelope (J. H. Carlson and T. J. Silhavy, J. Bacteriol. 175:3327-3334, 1993). We have characterized a multicopy suppressor of the processing-defective porins that appears to act by a novel mechanism. Using fractionation experiments and conformation-specific antibodies, we found that the presence of this multicopy suppressor allowed the processing-defective LamB precursors to be folded and localized to the outer membrane. Analysis of the suppressor plasmid revealed that these effects are mediated by the presence of a truncated derivative of the polytopic inner membrane protein, TetA. The suppression mediated by TetA' is independent of the CpxA/CpxR regulon and the sigma E regulon, both of which are involved in regulating protein trafficking functions in the cell envelope. |
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| Keywords |
Bacterial Outer Membrane Proteins; Base Sequence; Binding Sites/genetics; Cell Membrane/metabolism; DNA Primers/genetics; Escherichia coli/genetics; Escherichia coli/metabolism; Genes, Bacterial; Mutation; Porins; Protein Conformation; Protein Folding; Protein Precursors/chemistry; Protein Precursors/genetics; Protein Precursors/metabolism; Protein Processing, Post-Translational/genetics; Receptors, Virus/chemistry; Receptors, Virus/genetics; Receptors, Virus/metabolism; Suppression, Genetic |
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