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Induction of transient macroapertures in endothelial cells through RhoA inhibition by Staphylococcus aureus factors

¹ Toxines Bactériennes dans la Relation Hôte-Pathogènes, U627, and ² Biologie et Physiopathologie Cutanée, Faculté de Médecine, Institut National de la Santé et de la Recherche Médicale, INSERM, 06107 Nice Cedex 2, France
³ Centre Commun de Microscopie Électronique Appliquée, Faculté des Sciences, 06108 Nice Cedex 2, France
⁴ Laboratoire Central de Bactériologie, Hôpital ARCHET II, 06202 Nice Cedex 3, France
⁵ Unité des Interactions Bactéries-Cellules, Institut National de la Santé et de la Recherche Médicale, INSERM, U604, and ⁶ Unité des Bactéries Anaérobies et Toxines, Institut Pasteur, 75015 Paris, France

Correspondence to Emmanuel Lemichez: lemichez{at}unice.fr

The GTPase RhoA is a major regulator of the assembly of actin stress fibers and the contractility of the actomyosin cytoskeleton. The epidermal cell differentiation inhibitor (EDIN) and EDIN-like ADP-ribosyltransferases of Staphylococcus aureus catalyze the inactivation of RhoA, producing actin cable disruption. We report that purified recombinant EDIN and EDIN-producing S. aureus provoke large transcellular tunnels in endothelial cells that we have named macroapertures (MAs). These structures open transiently, followed by the appearance of actin-containing membrane waves extending over the aperture. Disruption of actin cables, either directly or indirectly, through rhoA RNAi knockdown also triggers the formation of MAs. Intoxication of endothelial monolayers by EDIN produces a loss of barrier function and provides direct access of the endothelium basement membrane to S. aureus.

Abbreviations used in this paper: EDIN, epidermal cell differentiation inhibitor; HMVEC, human microvascular endothelial cell; HUVEC, human umbilical vein endothelial cell; MA, macroaperture; RhoGDI, Rho guanine nucleotide dissociation inhibitor; ROCK, Rho kinase; SFM, serum-free medium; VVO, vesiculo-vacuolar organelle; WGA, wheat germ agglutinin.

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