Novel Topology of BfpE, a Cytoplasmic Membrane Protein Required for Type IV Fimbrial Biogenesis in Enteropathogenic Escherichia coli
10.1128/jb.183.15.4435-4450.2001
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

ABSTRACT Enteropathogenic Escherichia coli (EPEC) produces the bundle-forming pilus (BFP), a type IV fimbria that has been implicated in virulence, autoaggregation, and localized adherence to epithelial cells. The bfpE gene is one of a cluster of bfp genes previously shown to encode functions that direct BFP biosynthesis. Here, we show that an EPEC strain carrying a nonpolar mutation in bfpE fails to autoaggregate, adhere to HEp-2 cells, or form BFP, thereby demonstrating that BfpE is required for BFP biogenesis. BfpE is a cytoplasmic membrane protein of the GspF family. To determine the membrane topology of BfpE, we fused bfpE derivatives containing 3′ truncations and/or internal deletions to alkaline phosphatase and/or β-galactosidase reporter genes, whose products are active only when localized to the periplasm or cytoplasm, respectively. In addition, we constructed BfpE sandwich fusions using a dual alkaline phosphatase/β-galactosidase reporter cassette and analyzed BfpE deletion derivatives by sucrose density flotation gradient fractionation. The data from these analyses support a topology in which BfpE contains four hydrophobic transmembrane (TM) segments, a large cytoplasmic segment at its N terminus, and a large periplasmic segment near its C terminus. This topology is dramatically different from that of OutF, another member of the GspF family, which has three TM segments and is predominantly cytoplasmic. These findings provide a structural basis for predicting protein-protein interactions required for assembly of the BFP biogenesis machinery.

Bibliography

Blank, T. E., & Donnenberg, M. S. (2001). Novel Topology of BfpE, a Cytoplasmic Membrane Protein Required for Type IV Fimbrial Biogenesis in Enteropathogenic Escherichia coli. Journal of Bacteriology, 183(15), 4435–4450.

Authors 2
  1. T. Eric Blank (first)
  2. Michael S. Donnenberg (additional)
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Created 23 years, 1 month ago (July 27, 2002, 6:01 a.m.)
Deposited 4 years ago (July 29, 2021, 1:59 p.m.)
Indexed 6 hours, 45 minutes ago (Aug. 29, 2025, 5:57 a.m.)
Issued 24 years ago (Aug. 1, 2001)
Published 24 years ago (Aug. 1, 2001)
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@article{Blank_2001, title={Novel Topology of BfpE, a Cytoplasmic Membrane Protein Required for Type IV Fimbrial Biogenesis in Enteropathogenic Escherichia coli}, volume={183}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.183.15.4435-4450.2001}, DOI={10.1128/jb.183.15.4435-4450.2001}, number={15}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Blank, T. Eric and Donnenberg, Michael S.}, year={2001}, month=aug, pages={4435–4450} }