Identification of an actin‐binding site in p47<sup><i>phox</i></sup> an organizer protein of NADPH oxidase

Tamura, Minoru; Itoh, Katsunori; Akita, Hiroshi; Takano, Keiko; Oku, S.

doi:10.1016/j.febslet.2005.11.080

Cited by 24 publications

(15 citation statements)

References 31 publications

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“…In mammalian cells, the activity of NADPH oxidase is directly regulated by actin dynamics. Neutrophil NADPH oxidase binds to actin in vitro (Tamura et al, 2000a(Tamura et al, , 2000b(Tamura et al, , 2006. Actin polymerization activates NADPH oxidase, whereas depolymerizing actin deactivates it.…”

Section: Discussionmentioning

confidence: 99%

Capping Protein Modulates Actin Remodeling in Response to Reactive Oxygen Species during Plant Innate Immunity

Cao

Staiger

2016

Plant Physiol.

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Plants perceive microbe-associated molecular patterns and damage-associated molecular patterns to activate innate immune signaling events, such as bursts of reactive oxygen species (ROS). The actin cytoskeleton remodels during the first 5 min of innate immune signaling in Arabidopsis (Arabidopsis thaliana) epidermal cells; however, the immune signals that impinge on actin cytoskeleton and its response regulators remain largely unknown. Here, we demonstrate that rapid actin remodeling upon elicitation with diverse microbe-associated molecular patterns and damage-associated molecular patterns represent a conserved plant immune response. Actin remodeling requires ROS generated by the defense-associated NADPH oxidase, RBOHD. Moreover, perception of flg22 by its cognate receptor complex triggers actin remodeling through the activation of RBOHDdependent ROS production. Our genetic studies reveal that the ubiquitous heterodimeric capping protein transduces ROS signaling to the actin cytoskeleton during innate immunity. Additionally, we uncover a negative feedback loop between actin remodeling and flg22-induced ROS production.

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Section: Discussionmentioning

confidence: 99%

Capping Protein Modulates Actin Remodeling in Response to Reactive Oxygen Species during Plant Innate Immunity

Cao

Staiger

2016

Plant Physiol.

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“…phox (32). In vascular smooth muscle cells, the angiotensin II-mediated activation of NADPH oxidase and ROS generation is regulated in part by p47 phox /actin interaction through cortactin; however, the mechanisms of this interaction remain unclear (33).…”

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confidence: 99%

Regulation of Hyperoxia-induced NADPH Oxidase Activation in Human Lung Endothelial Cells by the Actin Cytoskeleton and Cortactin

Usatyuk

Romer

et al. 2007

Journal of Biological Chemistry

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Although the actin cytoskeleton has been implicated in the control of NADPH oxidase in phagocytosis, very little is known about the cytoskeletal regulation of endothelial NADPH oxidase assembly and activation. Here, we report a role for cortactin and the tyrosine phosphorylation of cortactin in hyperoxiainduced NADPH oxidase activation and ROS production in human pulmonary artery ECs (HPAECs). Exposure of HPAECs to hyperoxia for 3 h induced NADPH oxidase activation, as demonstrated by enhanced superoxide production. Hyperoxia also caused a thickening of the subcortical dense peripheral F-actin band and increased the localization of cortactin in the cortical regions and lamellipodia at cell-cell borders that protruded under neighboring cells. Pretreatment of HPAECs with the actin-stabilizing agent phallacidin attenuated hyperoxia-induced cortical actin thickening and ROS production, whereas cytochalasin D and latrunculin A enhanced basal and hyperoxia-induced ROS formation. In HPAECs, a 3-h hyperoxic exposure enhanced the tyrosine phosphorylation of cortactin and interaction between cortactin and p47 phox , a subcomponent of the EC NADPH oxidase, when compared with normoxic cells. Furthermore, transfection of HPAECs with cortactin small interfering RNA or myristoylated cortactin Src homology domain 3 blocking peptide attenuated ROS production and the hyperoxia-induced translocation of p47 phox to the cell periphery. Similarly, down-regulation of Src with Src small interfering RNA attenuated the hyperoxia-mediated phosphorylation of cortactin tyrosines and blocked the association of cortactin with actin and p47phox . In addition, the hyperoxia-induced generation of ROS was significantly lower in ECs expressing a tyrosine-deficient mutant of cortactin than in vector control or wild-type cells. These data demonstrate a novel function for cortactin and actin in hyperoxiainduced activation of NADPH oxidase and ROS generation in human lung endothelial cells. production that is dependent on NADPH oxidase activation and independent of the mitochondrial electron transport or xanthine/xanthine oxidase systems (10). The mechanisms of NADPH oxidase activation are complex. In phagocytes, activation of NADPH oxidase requires serine phosphorylation of the cytosolic p47 phox , p67 phox , and p40 phox components, assembly of the phosphorylated subunits with Rac2, and translocation to the phagosomes for association with cytochrome b 558. Here, one-electron reduction of molecular O 2 to O 2 . occurs with NADPH as the electron donor (7). In leukocytes, formyl-Met-Leu-Phe-OH or phorbol ester stimulates phosphorylation of p47 phox at multiple serine residues through reactions involving several protein kinases such as protein kinase C, protein kinase A, and mitogen-activated protein kinases (14 -17). In HPAECs, tumor necrosis factor-␣-medi-*This work was supported by National Institutes of Health Grants RO1 HL 69909 (to V. N.), PO1 HL 58064 (to V. N. and J. G. N. G.) and DE13079-01 and AI061042 (to L. H. R.). The costs of publication...

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“…For example, in a cell-free system, the addition of G-actin activated human neutrophil NADPH oxidase (31), whereas the addition of F-actin did not enhance oxidase activation. An actin binding site was identified in p47 phox (41). The present study showed that increased depolymerization of actin facilitated ROS production (Figs.…”

Section: Discussionmentioning

confidence: 68%

Endothelial actin depolymerization mediates NADPH oxidase-superoxide production during flow reversal

Choy

Yang

et al. 2014

American Journal of Physiology-Heart and Circulatory Physiology

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Slow moving blood flow and changes in flow direction, e.g., negative wall shear stress, can cause increased superoxide (O2 ·Ϫ ) production in vascular endothelial cells. The mechanism by which shear stress increases O2·Ϫ production, however, is not well established. We tested the hypothesis that actin depolymerization, which occurs during flow reversal, mediates O2 ·Ϫ production in vascular endothelial cells via NADPH oxidase, and more specifically, the subunit p47 phox . Using a swine model, we created complete blood flow reversal in one carotid artery, while the contralateral vessel maintained forward blood flow as control. We measured actin depolymerization, NADPH oxidase activity, and reactive oxygen species (ROS) production in the presence of various inhibitors. Flow reversal was found to induce actin depolymerization and a 3.9 Ϯ 1.0-fold increase in ROS production as compared with forward flow. NADPH oxidase activity was 1.4 Ϯ 0.2 times higher in vessel segments subjected to reversed blood flow when measured by a direct enzyme assay. The NADPH oxidase subunits gp91 phox (Nox2) and p47 phox content in the vessels remained unchanged after 4 h of flow reversal. In contrast, p47 phox phosphorylation was increased in vessels with reversed flow. The response caused by reversed flow was reduced by in vivo treatment with jasplakinolide, an actin stabilizer (only a 1.7 Ϯ 0.3-fold increase). Apocynin (an antioxidant) prevented reversed flow-induced ROS production when the animals were treated in vivo. Cytochalasin D mimicked actin depolymerization in vitro and caused a 5.2 Ϯ 3.0-fold increase in ROS production. These findings suggest that actin filaments play an important role in negative shear stress-induced ROS production by potentiating NADPH oxidase activity, and more specifically, the p47 phox subunit in vascular endothelium.endothelial cells

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Identification of an actin‐binding site in p47^phox an organizer protein of NADPH oxidase

Cited by 24 publications

References 31 publications

Capping Protein Modulates Actin Remodeling in Response to Reactive Oxygen Species during Plant Innate Immunity

Capping Protein Modulates Actin Remodeling in Response to Reactive Oxygen Species during Plant Innate Immunity

Regulation of Hyperoxia-induced NADPH Oxidase Activation in Human Lung Endothelial Cells by the Actin Cytoskeleton and Cortactin

Endothelial actin depolymerization mediates NADPH oxidase-superoxide production during flow reversal

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Identification of an actin‐binding site in p47phox an organizer protein of NADPH oxidase

Cited by 24 publications

References 31 publications

Capping Protein Modulates Actin Remodeling in Response to Reactive Oxygen Species during Plant Innate Immunity

Capping Protein Modulates Actin Remodeling in Response to Reactive Oxygen Species during Plant Innate Immunity

Regulation of Hyperoxia-induced NADPH Oxidase Activation in Human Lung Endothelial Cells by the Actin Cytoskeleton and Cortactin

Endothelial actin depolymerization mediates NADPH oxidase-superoxide production during flow reversal

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Identification of an actin‐binding site in p47^phox an organizer protein of NADPH oxidase