Abstract
Neisseria meningitidis is a Gram-negative diplococcus restricted to humans that causes severe septicemia and/or meningitidis. Initial adhesion to human endothelial cells is mediated through the interaction of type IV pili with the hetero-oligomeric complexes formed by the human receptors CD147 and the β2-adrenergic receptor. Interaction with this complex heterodimer activates a β-arrestin-biased signaling pathway leading to actin polymerization and accumulation of ezrin and ezrin-binding partners. These signaling events promote the formation of cell plasma membrane protrusions in endothelial cells, which are crucial for N. meningitidis colonies to resist shear stress and colonize blood vessels. Here we provide detailed protocols to evaluate the role of β-arrestins in actin and ezrin signaling downstream of G protein-coupled receptor activation.
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References
Join-Lambert O, Lecuyer H, Miller F, Lelievre L, Jamet A, Furio L, Schmitt A, Pelissier P, Fraitag S, Coureuil M, Nassif X (2013) Meningococcal interaction to microvasculature triggers the tissular lesions of purpura fulminans. J Infect Dis 208(10):1590–1597. https://doi.org/10.1093/infdis/jit301
Coureuil M, Lecuyer H, Bourdoulous S, Nassif X (2017) A journey into the brain: insight into how bacterial pathogens cross blood-brain barriers. Nat Rev Microbiol 15(3):149–159
Bernard SC, Simpson N, Join-Lambert O, Federici C, Laran-Chich MP, Maissa N, Bouzinba-Segard H, Morand PC, Chretien F, Taouji S, Chevet E, Janel S, Lafont F, Coureuil M, Segura A, Niedergang F, Marullo S, Couraud PO, Nassif X, Bourdoulous S (2014) Pathogenic Neisseria meningitidis utilizes CD147 for vascular colonization. Nat Med
Maissa N, Covarelli V, Janel S, Durel B, Simpson N, Bernard SC, Pardo-Lopez L, Bouzinba-Segard H, Faure C, Scott MGH, Coureuil M, Morand PC, Lafont F, Nassif X, Marullo S, Bourdoulous S (2017) Strength of Neisseria meningitidis binding to endothelial cells requires highly-ordered CD147/beta2-adrenoceptor clusters assembled by alpha-actinin-4. Nat Commun 8:15764
Coureuil M, Lecuyer H, Scott MG, Boularan C, Enslen H, Soyer M, Mikaty G, Bourdoulous S, Nassif X, Marullo S (2010) Meningococcus hijacks a beta2-adrenoceptor/beta-arrestin pathway to cross brain microvasculature endothelium. Cell 143(7):1149–1160
Nassif X, Lowy J, Stenberg P, O'Gaora P, Ganji A, So M (1993) Antigenic variation of pilin regulates adhesion of Neisseria meningitidis to human epithelial cells. Mol Microbiol 8(4):719–725
Weksler BB, Subileau EA, Perriere N, Charneau P, Holloway K, Leveque M, Tricoire-Leignel H, Nicotra A, Bourdoulous S, Turowski P, Male DK, Roux F, Greenwood J, Romero IA, Couraud PO (2005) Blood-brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J 19(13):1872–1874
Scott MG, Pierotti V, Storez H, Lindberg E, Thuret A, Muntaner O, Labbe-Jullie C, Pitcher JA, Marullo S (2006) Cooperative regulation of extracellular signal-regulated kinase activation and cell shape change by filamin A and beta-arrestins. Mol Cell Biol 26(9):3432–3445
Storez H, Scott MG, Issafras H, Burtey A, Benmerah A, Muntaner O, Piolot T, Tramier M, Coppey-Moisan M, Bouvier M, Labbe-Jullie C, Marullo S (2005) Homo- and hetero-oligomerization of beta-arrestins in living cells. J Biol Chem 280(48):40210–40215
Virji M, Watt SM, Barker S, Makepeace K, Doyonnas R (1996) The N-domain of the human CD66a adhesion molecule is a target for Opa proteins of Neisseria meningitidis and Neisseria gonorrhoeae. Mol Microbiol 22(5):929–939
Kellogg DS Jr, Cohen IR, Norins LC, Schroeter AL, Reising G (1968) Neisseria gonorrhoeae. II. Colonial variation and pathogenicity during 35 months in vitro. J Bacteriol 96(3):596–605
Luttrell LM, Ferguson SS, Daaka Y, Miller WE, Maudsley S, Della Rocca GJ, Lin F, Kawakatsu H, Owada K, Luttrell DK, Caron MG, Lefkowitz RJ (1999) Beta-arrestin-dependent formation of beta2 adrenergic receptor-Src protein kinase complexes. Science 283(5402):655–661
Soyer M, Charles-Orszag A, Lagache T, Machata S, Imhaus AF, Dumont A, Millien C, Olivo-Marin JC, Dumenil G (2014) Early sequence of events triggered by the interaction of Neisseria meningitidis with endothelial cells. Cell Microbiol 16(6):878–895
Billker O, Popp A, Gray-Owen SD, Meyer TF (2000) The structural basis of CEACAM-receptor targeting by neisserial Opa proteins. Trends Microbiol 8(6):258–260. discussion 260-251
Sanjana NE, Shalem O, Zhang F (2014) Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods 11(8):783–784
Merz AJ, Enns CA, So M (1999) Type IV pili of pathogenic Neisseriae elicit cortical plaque formation in epithelial cells. Mol Microbiol 32(6):1316–1332
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Virion, Z., Marullo, S., Coureuil, M. (2019). Methods to Study the Roles of β-Arrestins in Meningococcal Signaling. In: Scott, M., Laporte, S. (eds) Beta-Arrestins. Methods in Molecular Biology, vol 1957. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9158-7_20
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DOI: https://doi.org/10.1007/978-1-4939-9158-7_20
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