Lu/BCAM Adhesion Glycoprotein Is a Receptor for 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.


Vyšlo v časopise: Lu/BCAM Adhesion Glycoprotein Is a Receptor for Cytotoxic Necrotizing Factor 1 (CNF1). PLoS Pathog 10(1): e32767. doi:10.1371/journal.ppat.1003884
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003884

Souhrn

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.


Zdroje

1. UlettGC, TotsikaM, SchaaleK, CareyAJ, SweetMJ, et al. (2013) Uropathogenic Escherichia coli virulence and innate immune responses during urinary tract infection. Curr Opin Microbiol 16: S1369–5274.

2. PetkovsekZ, ElersicK, GubinaM, Zgur-BertokD, StarcicEM (2009) Virulence potential of Escherichia coli isolates from skin and soft tissue infections. J Clin Microbiol 47: 1811–1817.

3. KhanNA, WangY, KimKJ, ChungJW, WassCA, et al. (2002) Cytotoxic necrotizing factor-1 contributes to Escherichia coli K1 invasion of the central nervous system. J Biol Chem 277: 15607–15612.

4. FlatauG, LemichezE, GauthierM, ChardinP, ParisS, et al. (1997) Toxin-induced activation of the G protein p21 Rho by deamidation of glutamine. Nature 387: 729–733.

5. SchmidtG, SehrP, WilmM, SelzerJ, MannM, et al. (1997) Gln63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor 1. Nature 387: 725–729.

6. CherfilsJ, ZeghoufM (2013) Regulation of small GTPases by GEFs, GAPs, and GDIs. Physiol Rev 93: 269–309.

7. Schmidt G, Aktories K (2000) Rho GTPase-Activating Toxins: Cytotoxic Necrotizing Factors and Dermonecrotic Toxin. In: Balch WE, Der CJ, Hall A, editors. Regulators and Effectors of Small GTPases. San Diego: Academic Press. pp. 125–136.

8. ChungJW, HongSJ, KimKJ, GotiD, StinsMF, et al. (2003) 37 kDa laminin receptor precusor modulates cytotoxic necrotizing factor 1-mediated RhoA activation and bacterial uptake. J Biol Chem 278: 16857–16862.

9. NelsonJ, McFerranNV, PivatoG, ChambersE, DohertyC, et al. (2008) The 67 kDa laminin receptor: structure, function and role in disease. Biosci Rep 28: 33–48.

10. CollecE, ElNW, GauthierE, GaneP, LecomteMC, et al. (2007) Ubc9 interacts with Lu/BCAM adhesion glycoproteins and regulates their stability at the membrane of polarized MDCK cells. Biochem J 402: 311–319.

11. ElNW, GaneP, ColinY, D'AmbrosioAM, CallebautI, et al. (2001) Characterization of the laminin binding domains of the Lutheran blood group glycoprotein. J Biol Chem 276: 23757–23762.

12. BartolucciP, ChaarV, PicotJ, BachirD, HabibiA, et al. (2010) Decreased sickle red blood cell adhesion to laminin by hydroxyurea is associated with inhibition of Lu/BCAM protein phosphorylation. Blood 116: 2152–2159.

13. EylerCE, TelenMJ (2006) The Lutheran glycoprotein: a multifunctional adhesion receptor. Transfusion 46: 668–677.

14. KikkawaY, MinerJH (2005) Review: Lutheran/B-CAM: a laminin receptor on red blood cells and in various tissues. Connect Tissue Res 46: 193–199.

15. FatehullahA, DohertyC, PivatoG, AllenG, DevineL, et al. (2010) Interactions of the 67 kDa laminin receptor and its precursor with laminin. Biosci Rep 30: 73–79.

16. McNicholBA, RasmussenSB, CarvalhoHM, MeysickKC, O'BrienAD (2007) Two domains of cytotoxic necrotizing factor type 1 bind the cellular receptor, laminin receptor precursor protein. Infect Immun 75: 5095–5104.

17. BlumenthalB, HoffmannC, AktoriesK, BackertS, SchmidtG (2007) The Cytotoxic Necrotizing Factors from Yersinia pseudotuberculosis and from Escherichia coli Bind to Different Cellular Receptors but Take the Same Route to the Cytosol. Infect Immun 75: 3344–3353.

18. RahuelC, FilipeA, RitieL, ElNW, Patey-MariaudN, et al. (2008) Genetic inactivation of the laminin alpha5 chain receptor Lu/BCAM leads to kidney and intestinal abnormalities in the mouse. Am J Physiol Renal Physiol 294: F393–F406.

19. ChenC, FuZ, KimJJ, BarbieriJT, BaldwinMR (2009) Gangliosides as high affinity receptors for tetanus neurotoxin. J Biol Chem 284: 26569–26577.

20. StollT, MarkwirthG, ReipschlagerS, SchmidtG (2009) A new member of a growing toxin family - Escherichia coli cytotoxic necrotizing factor 3 (CNF3). Toxicon 54: 745–753.

21. LemichezE, FlatauG, BruzzoneM, BoquetP, GauthierM (1997) Molecular localization of the Escherichia coli cytotoxic necrotizing factor CNF1 cell-binding and catalytic domains. Mol Microbiol 24: 1061–1070.

22. PeiS, DoyeA, BoquetP (2001) Mutation of specific acidic residues of the CNF1 T domain into lysine alters cell membrane translocation of the toxin. Mol Microbiol 41: 1237–1247.

23. HoffmannC, SchmidtG (2004) CNF and DNT. Rev Physiol Biochem Pharmacol 152: 49–63.

24. GerhardR, SchmidtG, HofmannF, AktoriesK (1998) Activation of Rho GTPases by Escherichia coli cytotoxic necrotizing factor 1 increases intestinal permeability in Caco-2 cells. Infect Immun 66: 5125–5131.

25. WilkeGA, BubeckWJ (2010) Role of a disintegrin and metalloprotease 10 in Staphylococcus aureus alpha-hemolysin-mediated cellular injury. Proc Natl Acad Sci U S A 107: 13473–13478.

26. KimKJ, ChungJW, KimKS (2005) 67-kDa laminin receptor promotes internalization of cytotoxic necrotizing factor 1-expressing Escherichia coli K1 into human brain microvascular endothelial cells. J Biol Chem 280: 1360–1368.

27. RattsR, ZengH, BergEA, BlueC, McCombME, et al. (2003) The cytosolic entry of diphtheria toxin catalytic domain requires a host cell cytosolic translocation factor complex. J Cell Biol 160: 1139–1150.

28. BuetowL, FlatauG, ChiuK, BoquetP, GhoshP (2001) Structure of the Rho-activating domain of Escherichia coli cytotoxic necrotizing factor 1. Nature Struct Biol 8: 584–588.

29. KnustZ, BlumenthalB, AktoriesK, SchmidtG (2009) Cleavage of Escherichia coli cytotoxic necrotizing factor 1 is required for full biologic activity. Infect Immun 77: 1835–1841.

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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