Arg188 drives RhoC membrane binding

Patel, Aditi; Williams-Perez, Sophia; Peyton, Nicole; Reicks, Amy; Buzick, Justin; Farley, Janean; Shirar, Sarah; Ellerbroek, Shawn M.

doi:10.1080/21541248.2016.1205334

Cited by 6 publications

(5 citation statements)

References 25 publications

(31 reference statements)

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“…Further studies should identify the specific GEFs and GAPs for RhoC to better understand the molecular details of the signal transduction pathway during phagosome formation. Recently, Patel et al have reported that RhoC shows a higher degree of membrane association than does RhoA (Patel et al, 2016). In their study, these authors demonstrated that arginine 188 of RhoC promotes membrane binding.…”

Section: Discussionmentioning

confidence: 94%

RhoC regulates the actin remodeling required for phagosome formation during FcγR-mediated phagocytosis

Egami

Kawai

Araki

2017

Journal of Cell Science

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Phagosome formation is a complicated process that requires spatiotemporally regulated actin reorganization. We found that RhoC GTPase is a critical regulator of FcγR-mediated phagocytosis in macrophages. Our live-cell imaging revealed that RhoC, but not RhoA, is recruited to phagocytic cups engulfing IgG-opsonized erythrocytes (IgG-Es). RhoC silencing through RNAi, CRISPR/Cas-mediated RhoC knockout, and the expression of dominant-negative or constitutively active RhoC mutants suppressed the phagocytosis of IgG-Es. Moreover, RhoC-GTP pulldown experiments showed that endogenous RhoC is transiently activated during phagosome formation. Notably, actin-driven pseudopod extension, which is required for the formation of phagocytic cups, was severely impaired in cells expressing the constitutively active mutant RhoC-G14V, which induced abnormal F-actin accumulation underneath the plasma membrane. mDia1 (encoded by ), a Rho-dependent actin nucleation factor, and RhoC were colocalized at the phagocytic cups. Similar to what was seen for RhoC, mDia1 silencing through RNAi inhibited phagosome formation. Additionally, the coexpression of mDia1 with constitutively active mutant RhoC-G14V or expression of active mutant mDia1-ΔN3 drastically inhibited the uptake of IgG-Es. These data suggest that RhoC modulates phagosome formation be modifying actin cytoskeletal remodeling via mDia1.

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Section: Discussionmentioning

confidence: 94%

RhoC regulates the actin remodeling required for phagosome formation during FcγR-mediated phagocytosis

Egami

Kawai

Araki

2017

Journal of Cell Science

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“…31 Within the RhoA/ROCK signaling pathway, RhoA phosphorylation at Ser188 directly inhibits RhoA activity, aligning with the findings of our study. 32,33 Moreover, this phosphorylation of RhoA serves as a key protective mechanism against RhoAmediated vascular remodeling. 34 In this study, we first investigated whether H 2 S could promote RhoA phosphorylation at Ser188, thereby inhibiting the activity of the RhoA/ROCK pathway in vivo.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Hydrogen Sulfide Protects against Rat Ischemic Brain Injury by Promoting RhoA Phosphorylation at Serine 188

Chen,

et al. 2024

ACS Omega

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The protective role of hydrogen sulfide against cerebral ischemia-reperfusion injury involves the inhibition of the RhoA-/Rho-associated coiled-coil kinase (ROCK) pathway. However, the specific mechanism remains elusive. This study investigates the impact of hydrogen sulfide on RhoA phosphorylation at serine 188 (Ser188) in vivo, aiming to test the hypothesis that hydrogen sulfide exerts neuroprotection by enhancing RhoA phosphorylation at Ser188, subsequently inhibiting the RhoA/ROCK pathway. Recombinant RhoA wild -pEGFP-N1 and RhoA S188A -pEGFP-N1 plasmids were constructed and administered via stereotaxic injection into the rat hippocampus. A rat global cerebral ischemia-reperfusion model was induced by bilateral carotid artery ligation to elucidate the neuroprotective mechanisms of hydrogen sulfide. Both RhoA wild -pEGFP-N1 and RhoA S188A -pEGFP-N1 plasmids expressed RhoA wild and RhoA S188A proteins, respectively, in rat hippocampal tissues, alongside the intrinsic RhoA protein. Systemic administration of the exogenous hydrogen sulfide donor sodium hydrosulfide led to an increase in Ser188 phosphorylation of transfected RhoA wild and intrinsic RhoA protein within the hippocampus. However, this effect was not observed in tissues transfected with RhoA S188A . Sodium hydrosulfide-mediated RhoA phosphorylation correlated with decreased RhoA and ROCK 2 activity in rat hippocampal tissues. Furthermore, sodium hydrosulfide administration reduced cerebral ischemia-reperfusion-induced neuronal damage and apoptosis in rat hippocampal tissues transfected with RhoA wild . However, this neuroprotective effect was attenuated in rats transfected with RhoA S188A . These findings suggest that the neuroprotective mechanism of hydrogen sulfide against cerebral ischemia/reperfusion injury involves increased RhoA phosphorylation at Ser188. Promoting this phosphorylation may represent a potential intrinsic therapeutic target for ischemic stroke.

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“…It is widely recognized that phosphorylation of RhoA at Ser188 directly inhibits RhoA activity 35 , but phosphorylation at Ser88 increases its activity 36 . In the present study, we demonstrated that RhoA activity is significantly inhibited in HNCs after phosphorylation at Ser188, which is consistent with previous results 37 .…”

Section: Discussionmentioning

confidence: 99%

H2S protects hippocampal neurons against hypoxia-reoxygenation injury by promoting RhoA phosphorylation at Ser188

2021

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Inhibition of RhoA-ROCK pathway is involved in the H2S-induced cerebral vasodilatation and H2S-mediated protection on endothelial cells against oxygen-glucose deprivation/reoxygenation injury. However, the inhibitory mechanism of H2S on RhoA-ROCK pathway is still unclear. The aim of this study was to investigate the target and mechanism of H2S in inhibition of RhoA/ROCK. GST-RhoAwild and GST-RhoAS188A proteins were constructed and expressed, and were used for phosphorylation assay in vitro. Recombinant RhoAwild-pEGFP-N1 and RhoAS188A-pEGFP-N1 plasmids were constructed and transfected into primary hippocampal nerve cells (HNCs) to evaluate the neuroprotective mechanism of endothelial H2S by using transwell co-culture system with endothelial cells from cystathionine-γ-lyase knockout (CSE−/−) mice and 3-mercaptopyruvate sulfurtransferase knockout (3-MST−/−) rats, respectively. We found that NaHS, exogenous H2S donor, promoted RhoA phosphorylation at Ser188 in the presence of cGMP-dependent protein kinase 1 (PKG1) in vitro. Besides, both exogenous and endothelial H2S facilitated the RhoA phosphorylation at Ser188 in HNCs, which induced the reduction of RhoA activity and membrane transposition, as well as ROCK2 activity and expression. To further investigate the role of endothelial H2S on RhoA phosphorylation, we detected H2S release from ECs of CSE+/+ and CSE−/− mice, and 3-MST+/+ and 3-MST−/− rats, respectively, and found that H2S produced by ECs in the culture medium is mainly catalyzed by CSE synthase. Moreover, we revealed that both endothelial H2S, mainly catalyzed by CSE, and exogenous H2S protected the HNCs against hypoxia-reoxygenation injury via phosphorylating RhoA at Ser188.

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Arg188 drives RhoC membrane binding

Cited by 6 publications

References 25 publications

RhoC regulates the actin remodeling required for phagosome formation during FcγR-mediated phagocytosis

RhoC regulates the actin remodeling required for phagosome formation during FcγR-mediated phagocytosis

Hydrogen Sulfide Protects against Rat Ischemic Brain Injury by Promoting RhoA Phosphorylation at Serine 188

H2S protects hippocampal neurons against hypoxia-reoxygenation injury by promoting RhoA phosphorylation at Ser188

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