Correction: Lu/BCAM Adhesion Glycoprotein Is a Receptor for Escherichia coli Cytotoxic Necrotizing Factor 1 (CNF1)

The Cytotoxic Necrotizing Factor 1 (CNF1) is a protein toxin which is a major virulence factor of pathogenic Escherichia coli strains. Here, we identified the Lutheran (Lu) adhesion glycoprotein/basal cell adhesion molecule (BCAM) as cellular receptor for CNF1 by co-precipitation of cell surface molecules with tagged toxin. The CNF1-Lu/BCAM interaction was verified by direct protein-protein interaction analysis and competition studies. These studies revealed amino acids 720 to 1014 of CNF1 as the binding site for Lu/BCAM. We suggest two cell interaction sites in CNF1: first the N-terminus, which binds to p37LRP as postulated before. Binding of CNF1 to p37LRP seems to be crucial for the toxin's action. However, it is not sufficient for the binding of CNF1 to the cell surface. A region directly adjacent to the catalytic domain is a high affinity interaction site for Lu/BCAM. We found Lu/BCAM to be essential for the binding of CNF1 to cells. Cells deficient in Lu/BCAM but expressing p37LRP could not bind labeled CNF1. Therefore, we conclude that LRP and Lu/BCAM are both required for toxin action but with different functions. Copyright: ß 2014 Piteau et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

[1]  M. Sweet,et al.  Uropathogenic Escherichia coli virulence and innate immune responses during urinary tract infection. , 2013, Current opinion in microbiology.

[2]  Jacqueline Cherfils,et al.  Regulation of small GTPases by GEFs, GAPs, and GDIs. , 2013, Physiological reviews.

[3]  D. Bachir,et al.  Decreased sickle red blood cell adhesion to laminin by hydroxyurea is associated with inhibition of Lu/BCAM protein phosphorylation. , 2010, Blood.

[4]  J. B. Wardenburg,et al.  Role of a disintegrin and metalloprotease 10 in Staphylococcus aureus α-hemolysin–mediated cellular injury , 2010, Proceedings of the National Academy of Sciences.

[5]  Simone Reipschläger,et al.  A new member of a growing toxin family--Escherichia coli cytotoxic necrotizing factor 3 (CNF3). , 2009, Toxicon : official journal of the International Society on Toxinology.

[6]  C. Doherty,et al.  Interactions of the 67 kDa laminin receptor and its precursor with laminin , 2009, Bioscience reports.

[7]  J. Barbieri,et al.  Gangliosides as High Affinity Receptors for Tetanus Neurotoxin* , 2009, The Journal of Biological Chemistry.

[8]  D. Žgur-Bertok,et al.  Virulence Potential of Escherichia coli Isolates from Skin and Soft Tissue Infections , 2009, Journal of Clinical Microbiology.

[9]  K. Aktories,et al.  Cleavage of Escherichia coli Cytotoxic Necrotizing Factor 1 Is Required for Full Biologic Activity , 2009, Infection and Immunity.

[10]  J. Cartron,et al.  Genetic inactivation of the laminin alpha5 chain receptor Lu/BCAM leads to kidney and intestinal abnormalities in the mouse. , 2008, American journal of physiology. Renal physiology.

[11]  C. Doherty,et al.  The 67 kDa laminin receptor: structure, function and role in disease. , 2008, Bioscience reports.

[12]  A. O’Brien,et al.  Two Domains of Cytotoxic Necrotizing Factor Type 1 Bind the Cellular Receptor, Laminin Receptor Precursor Protein , 2007, Infection and Immunity.

[13]  S. Backert,et al.  The Cytotoxic Necrotizing Factors from Yersinia pseudotuberculosis and from Escherichia coli Bind to Different Cellular Receptors but Take the Same Route to the Cytosol , 2007, Infection and Immunity.

[14]  P. Gane,et al.  Ubc9 interacts with Lu/BCAM adhesion glycoproteins and regulates their stability at the membrane of polarized MDCK cells. , 2007, The Biochemical journal.

[15]  M. Telen,et al.  The Lutheran glycoprotein: a multifunctional adhesion receptor , 2006, Transfusion.

[16]  J. Chung,et al.  67-kDa Laminin Receptor Promotes Internalization of Cytotoxic Necrotizing Factor 1-expressing Escherichia coli K1 into Human Brain Microvascular Endothelial Cells* , 2005, Journal of Biological Chemistry.

[17]  Y. Kikkawa,et al.  Review: Lutheran/B-CAM: A Laminin Receptor on Red Blood Cells and in Various Tissues , 2005, Connective tissue research.

[18]  T. Dawson,et al.  37-kDa Laminin Receptor Precursor Modulates Cytotoxic Necrotizing Factor 1–mediated RhoA Activation and Bacterial Uptake* , 2003, The Journal of Biological Chemistry.

[19]  M. McComb,et al.  The cytosolic entry of diphtheria toxin catalytic domain requires a host cell cytosolic translocation factor complex , 2003, Journal of Cell Biology.

[20]  J. Chung,et al.  Cytotoxic Necrotizing Factor-1 Contributes to Escherichia coli K1 Invasion of the Central Nervous System* , 2002, The Journal of Biological Chemistry.

[21]  P. Boquet,et al.  Mutation of specific acidic residues of the CNF1 T domain into lysine alters cell membrane translocation of the toxin , 2001, Molecular microbiology.

[22]  P. Boquet,et al.  Structure of the Rho-activating domain of Escherichia coli cytotoxic necrotizing factor 1 , 2001, Nature Structural Biology.

[23]  Isabelle Callebaut,et al.  Characterization of the Laminin Binding Domains of the Lutheran Blood Group Glycoprotein* , 2001, The Journal of Biological Chemistry.

[24]  K. Aktories,et al.  Rho GTPase-activating toxins: cytotoxic necrotizing factors and dermonecrotic toxin. , 2000, Methods in enzymology.

[25]  K. Aktories,et al.  Activation of Rho GTPases by Escherichia coli Cytotoxic Necrotizing Factor 1 Increases Intestinal Permeability in Caco-2 Cells , 1998, Infection and Immunity.

[26]  C. Fiorentini,et al.  Toxin-induced activation of the G protein p21 Rho by deamidation of glutamine , 1997, Nature.

[27]  M. Mann,et al.  Gln 63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor-1 , 1997, Nature.

[28]  P. Boquet,et al.  Molecular localization of the Escherichia coli cytotoxic necrotizing factor CNF1 cell‐binding and catalytic domains , 1997, Molecular microbiology.