Type IV pilus secretins have extracellular C termini

Joshua A. Lieberman, Courtney D. Petro, Stefani Thomas, Austin Yang, Michael S. Donnenberg

Research output: Contribution to journalArticle

Abstract

Type IV pili (T4Ps) are surface appendages used by Gram-negative and Gram-positive pathogens for motility and attachment to epithelial surfaces. In Gram-negative bacteria, such as the important pediatric pathogen enteropathogenic Escherichia coli (EPEC), during extension and retraction, the pilus passes through an outer membrane (OM) pore formed by the multimeric secretin complex. The secretin is common to Gram-negative assemblies, including the related type 2 secretion (T2S) system and the type 3 secretion (T3S) system. The N termini of the secretin monomers are periplasmic and in some systems have been shown to mediate substrate specificity. In this study, we mapped the topology of BfpB, the T4P secretin from EPEC, using a combination of biochemical and biophysical techniques that allowed selective identification of periplasmic and extracellular residues. We applied rules based on solved atomic structures of outer membrane proteins (OMPs) to generate our topology model, combining the experimental results with secondary structure prediction algorithms and direct inspection of the primary sequence. Surprisingly, the C terminus of BfpB is extracellular, a result confirmed by flow cytometry for BfpB and a distantly related T4P secretin, PilQ, from Pseudomonas aeruginosa. Keeping with prior evidence, the C termini of two T2S secretins and one T3S secretin were not detected on the extracellular surface. On the basis of our data and structural constraints, we propose that BfpB forms a beta barrel with 16 transmembrane beta strands. We propose that the T4P secretins have a C-terminal segment that passes through the center of each monomer.

Original languageEnglish (US)
Article numbere00322-15
JournalmBio
Volume6
Issue number2
DOIs
StatePublished - Mar 24 2015

ASJC Scopus subject areas

  • Microbiology
  • Virology

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  • Cite this

    Lieberman, J. A., Petro, C. D., Thomas, S., Yang, A., & Donnenberg, M. S. (2015). Type IV pilus secretins have extracellular C termini. mBio, 6(2), [e00322-15]. https://doi.org/10.1128/mBio.00322-15