Reference : The recruitment of p47 and Rac2G12V at the phagosome is transient and phosphatidylser...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
The recruitment of p47 and Rac2G12V at the phagosome is transient and phosphatidylserine dependent.
Faure, Marie Cecile [> >]
Sulpice, Jean-Claude [> >]
Delattre, Maud [> >]
Lavielle, Marc [> >]
Prigent, Magali [> >]
Cuif, Marie-Helene [> >]
Melchior, Chantal [> >]
Tschirhart, Eric mailto [University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit >]
Nusse, Oliver [> >]
Dupre-Crochet, Sophie [> >]
Biology of the Cell
Yes (verified by ORBilu)
[en] Anionic phospholipids ; Live imaging ; NADPH oxidase ; Phagocytosis ; ROS production
[en] BACKGROUND INFORMATION: During phagocytosis, neutrophils internalise pathogens in a phagosome and produce reactive oxygen species (ROS) by the NADPH oxidase to kill the pathogen. The cytosolic NADPH oxidase subunits p40phox , p47phox , p67phox and Rac2 translocate to the phagosomal membrane to participate in enzyme activation. The kinetics of this recruitment and the underlying signalling pathways are only partially understood. Anionic phospholipids, phosphatidylserine (PS) and phosphoinositides (PPI) provide an important attachment site for numerous proteins, including several oxidase subunits. RESULTS: We investigated the kinetics of p47phox and Rac2 phagosomal membrane recruitment. Both subunits are known to interact with anionic phospholipids; we therefore addressed the role of PS in this recruitment. Phagosomal accumulation of p47phox and Rac2 tagged with fluorescent proteins was analysed by videomicroscopy. We used the C2 domain of lactadherin (lactC2) that interacts strongly and specifically with PS to monitor intracellular PS localisation and to decrease PS accessibility. During phagocytosis of opsonised zymosan, p47phox and constitutively active Rac2G12V briefly translocated to the phagosomal membrane, whereas ROS production continued for a longer period. However, in the presence of lactC2, Rac2G12V recruitment was inhibited and the kinetics of p47phox recruitment and detachment were delayed. A reduced phagosomal ROS production was also observed during the first 7 min following the phagosome closure. CONCLUSIONS: These results suggest that p47phox and Rac2 accumulate only transiently at the phagosome at the onset of NADPH activity and detach from the phagosome before the end of ROS production. Furthermore, lactC2, by masking PS, interfered with the phagosomal recruitment of p47phox and Rac2 and disturbed NADPH oxidase activity. Thus, PS appears as a modulator of NADPH oxidase activation.
(c) 2013 Societe Francaise des Microscopies and Societe de Biologie Cellulaire de France.

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