title: An unusual signal peptide facilitates late steps in the biogenesis of a bacterial autotransporter.
abstract: Bacterial autotransporters are proteins that use a C-terminal porin-like domain to facilitate the transport of an upstream "passenger domain" across the outer membrane. Although autotransporters are translocated across the inner membrane (IM) via the Sec pathway, some of them contain exceptionally long signal peptides distinguished by a unique N-terminal sequence motif. In this study, we used the Escherichia coli O157:H7 autotransporter EspP as a model protein to investigate the function of the unusual signal peptides. We found that removal of the N-terminal motif or replacement of the EspP signal peptide did not affect translocation of the protein across the IM. Remarkably, modification of the signal peptide caused EspP to misfold in the periplasm and blocked transport of the passenger domain across the outer membrane. Further analysis suggested that the EspP signal peptide transits slowly through the Sec machinery. Based on these results, we propose that the unusual signal peptides not only function as targeting signals, but also prevent misfolding of the passenger domain in the periplasm by transiently tethering it to the IM.
authors.affiliation: Genetics and Biochemistry Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Rose L
authors.initials: RL
authors.lastname: Szabady
authors.forename: Janine H
authors.initials: JH
authors.lastname: Peterson
authors.forename: Kristen M
authors.initials: KM
authors.lastname: Skillman
authors.forename: Harris D
authors.initials: HD
authors.lastname: Bernstein
mesh_terms: D004926:Escherichia coli / Q000502:physiology; D029968:Escherichia coli Proteins / Q000096:biosynthesis*; D011499:Protein Processing, Post-Translational / Q000502:physiology; D021382:Protein Sorting Signals / Q000502:physiology*; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000502:physiology; D012697:Serine Endopeptidases / Q000096:biosynthesis*
chemical_list: D029968:Escherichia coli Proteins; D021382:Protein Sorting Signals; C107189:EspP protein, E coli; D012697:Serine Endopeptidases