CBL1‐CIPK26‐mediated phosphorylation enhances activity of the NADPH oxidase RBOHC, but is dispensable for root hair growth

Zhang, Xinxin; Köster, Philipp; Schlücking, Kathrin; Balcerowicz, Daria; Hashimoto, Kenji; Kuchitsu, Kazuyuki; Vissenberg, Kris; Kudla, Jörg

doi:10.1002/1873-3468.13187

Cited by 32 publications

(21 citation statements)

References 49 publications

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“…64,65 Serine phosphorylation can occur through the interaction of RBOHC and the calcineurin B-like-interacting protein kinase 26 (CIPK26)-calcineurin B-like protein 1 (CBL1) sensor complex. 47,66 Heterologous expression of wild-type or serine-to-alanine mutated versions of RBOHC showed that ROS production was dampened in serine-mutated RBOHC proteins, suggesting that phosphorylation at S318 and S322 amplifies ROS production in response to Ca 2+ influx. 66 The authors suggested that phosphorylation of RBOHC may act in concert with Ca 2+ oscillation to modulate ROS production in developing root hairs.…”

Section: Primary Root Development Andmentioning

confidence: 99%

“…47,66 Heterologous expression of wild-type or serine-to-alanine mutated versions of RBOHC showed that ROS production was dampened in serine-mutated RBOHC proteins, suggesting that phosphorylation at S318 and S322 amplifies ROS production in response to Ca 2+ influx. 66 The authors suggested that phosphorylation of RBOHC may act in concert with Ca 2+ oscillation to modulate ROS production in developing root hairs. 49,66 Interestingly, root hair development in cbl1/cipk26 double mutants and single mutants and in rhd2 mutant lines harboring the serine-mutated RBOHC protein displayed wild-type root hair morphology, suggesting that the two conserved serine residues are not required in planta or that other phosphorylation sites/kinases are involved in RBOHC activity.…”

Section: Primary Root Development Andmentioning

confidence: 99%

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RBOH-Dependent ROS Synthesis and ROS Scavenging by Plant Specialized Metabolites To Modulate Plant Development and Stress Responses

Chapman

Mühlemann

Gayomba

et al. 2019

Chem. Res. Toxicol.

218

162

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Reactive oxygen species (ROS) regulate plant growth and development. ROS are kept at low levels in cells to prevent oxidative damage, allowing them to be effective signaling molecules upon increased synthesis. In plants and animals, NADPH oxidase/respiratory burst oxidase homolog (RBOH) proteins provide localized ROS bursts to regulate growth, developmental processes, and stress responses. This review details ROS production via RBOH enzymes in the context of plant development and stress responses and defines the locations and tissues in which members of this family function in the model plant Arabidopsis thaliana. To ensure that these ROS signals do not reach damaging levels, plants use an array of antioxidant strategies. In addition to antioxidant machineries similar to those found in animals, plants also have a variety of specialized metabolites that scavenge ROS. These plant specialized metabolites exhibit immense structural diversity and have highly localized accumulation. This makes them important players in plant developmental processes and stress responses that use ROS-dependent signaling mechanisms. This review summarizes the unique properties of plant specialized metabolites, including carotenoids, ascorbate, tocochromanols (vitamin E), and flavonoids, in modulating ROS homeostasis. Flavonols, a subclass of flavonoids with potent antioxidant activity, are induced during stress and development, suggesting that they have a role in maintaining ROS homeostasis. Recent results using genetic approaches have shown how flavonols regulate development and stress responses through their action as antioxidants.

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Section: Primary Root Development Andmentioning

confidence: 99%

Section: Primary Root Development Andmentioning

confidence: 99%

RBOH-Dependent ROS Synthesis and ROS Scavenging by Plant Specialized Metabolites To Modulate Plant Development and Stress Responses

Chapman

Mühlemann

Gayomba

et al. 2019

Chem. Res. Toxicol.

218

162

View full text Add to dashboard Cite

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“…To measure the activity of RBOHF, pEF1-2xStrepII-N or pEF1-2xStrepII-C vectors (Drerup et al, 2013) containing RBOHF or other gene coding sequences (CBL1, CIPK3, CIPK11, CIPK26, OST1, ABI1) were transiently transfected into HEK293T cells, and the ROS production assay was performed as described previously (Zhang et al, 2018). In each assay, 10 min after starting measurements, 1 lM ionomycin and 1 mM CaCl 2 were automatically injected into the wells to induce Ca 2+ influx into the cells.…”

Section: Rbohf Activity Measurements In Hek293t Cellsmentioning

confidence: 99%

“…Among the protein modifications affecting RBOH activity, protein phosphorylation has been emerging as playing a key role in NADPH oxidase regulation. Genetic and biochemical studies suggested that Ca 2+ -dependent phosphorylation is required for RBOHC-mediated regulation of root hair growth and, recently, the Ca 2+ -activated calcineurin B-like (CBL) interacting protein kinase (CIPK) CIPK26 has been identified as directly phosphorylating RBOHC (Takeda et al, 2008;Zhang et al, 2018). Also, RBOHD is subject to regulation by phosphorylation.…”

Section: Introductionmentioning

confidence: 99%

Fine‐tuning of RBOHF activity is achieved by differential phosphorylation and Ca²⁺ binding

et al. 2018

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Summary RBOHF from Arabidopsis thaliana represents a multifunctional NADPH oxidase regulating biotic and abiotic stress tolerance, developmental processes and guard cell aperture. The molecular components and mechanisms determining RBOHF activity remain to be elucidated. Here we combined protein interaction studies, biochemical and genetic approaches, and pathway reconstitution analyses to identify and characterize proteins that confer RBOHF regulation and elucidated mechanisms that adjust RBOHF activity. While the Ca2+ sensor‐activated kinases CIPK11 and CIPK26 constitute alternative paths for RBOHF activation, the combined activity of CIPKs and the kinase open stomata 1 (OST1) triggers complementary activation of this NADPH oxidase, which is efficiently counteracted through dephosphorylation by the phosphatase ABI1. Within RBOHF, several distinct phosphorylation sites (p‐sites) in the N‐terminus of RBOHF appear to contribute individually to activity regulation. These findings identify RBOHF as a convergence point targeted by a complex regulatory network of kinases and phosphatases. We propose that this allows for fine‐tuning of plant reactive oxygen species (ROS) production by RBOHF in response to different stimuli and in diverse physiological processes.

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“…Transformation and cultivation of HEK293T cells was performed as described previously (Ogasawara et al, 2008; Zhang et al, 2018). Spectral characteristics of ABA indicators were recorded in Greiner flat bottom 96-well plates (Greiner BIO-ONE) using a TECAN Safire plate reader (TECAN) operated by the XFLUOR4.51 software with the following parameters: fluorescence emission scan bottom mode; excitation wavelength 440 nm, bandwidth 12.5 nm; emission wavelength scan from 460-600 nm, bandwidth 10 nm; gain 100-115; flashes 10; integration time 40-60 µs; temperature 37 °C.…”

Section: Methodsmentioning

confidence: 99%

Dual-sensing genetically encoded fluorescent indicators resolve the spatiotemporal coordination of cytosolic abscisic acid and second messenger dynamics in Arabidopsis

Waadt

Köster

Andrés

et al. 2019

Preprint

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Dual-sensing genetically encoded fluorescent indicators resolve the 22 spatiotemporal coordination of cytosolic abscisic acid and second messenger 23 dynamics in Arabidopsis 24 SHORT TITLE 25 Coordination of signaling compound fluxes in roots 26 Rainer Waadt et al. -Coordination of signaling compound fluxes in roots 2 27 ABSTRACT 28 Deciphering signal transduction processes is crucial for understanding 29 how plants sense and respond to environmental changes. Various chemical 30 compounds function as central messengers within deeply intertwined signaling 31 networks. How such compounds act in concert remains to be elucidated. We 32 have developed dual-sensing genetically encoded fluorescent indicators (2-In-1-33 GEFIs) for multiparametric in vivo analyses of the phytohormone abscisic acid 34 (ABA), Ca 2+ , protons (H + ), chloride (anions), the glutathione redox potential (E GSH ) 35 and hydrogen peroxide (H 2 O 2 ). Simultaneous analyses of two signaling 36 compounds in Arabidopsis (Arabidopsis thaliana) roots revealed that ABA 37 treatment and uptake did not trigger rapid cytosolic Ca 2+ or H + fluxes. Glutamate, 38 ATP, Arabidopsis PLANT ELICITOR PEPTIDE (AtPEP1) and glutathione disulfide 39 (GSSG) treatments induced rapid spatiotemporally overlapping cytosolic Ca 2+ , H + 40 and anion fluxes, but except for GSSG only weakly affected the cytosolic redox 41 state. Overall, 2-In-1-GEFIs enable complementary high-resolution in vivo 42 analyses of signaling compound dynamics and facilitate an advanced 43 understanding of the spatiotemporal coordination of signal transduction 44 processes in Arabidopsis. 45 46 198 Supplemental Movie 3). These experiments established the functionality of both 2-In-1-199 GEFIs and revealed that ABA does not trigger rapid cytosolic Ca 2+ or pH changes in 200 roots. 201 Auxin, ATP and glutamate treatments trigger spatiotemporally overlapping fluxes 202 of Ca 2+ , H + and Cl -203 Rainer Waadt et al. -Coordination of signaling compound fluxes in roots 8 Next, we used R-GECO1-GSL-E 2 GFP to simultaneously monitor Ca 2+ , H + and Cl -204 fluxes. Because anions, such as Cl -, quench the fluorescence of E 2 GFP and because its 205 excitation ratiometric pH readout is Clindependent, E 2 GFP provides a means to 206 simultaneously assess cytosolic H + and Clchanges (Bizzarri et al., 2006; Arosio et al., 207 2010). In response to 1 µM IAA, R-GECO1 reported biphasic Ca 2+ signals in the root 208 elongation zone that travelled to neighboring regions. Subsequent 1 mM ATP 209 treatments triggered Ca 2+ signals in the calyptra and meristematic zone that proceeded 210 shootward (Figure 4A left). Interestingly, both Ca 2+ signals coincided with a cytosolic 211 acidification reported by E 2 GFP (Figure 4A middle, Supplemental Movie 4). IAA also 212 induced a Clinflux, indicated by a E 2 GFP fluorescence emission decrease in the entire 213 imaged root, with a maximum decrease in the meristematic zone (Figure 4A right). ATP 214 however induced Clinflux in the upper elongation zone and above, but Clefflux ...

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CBL1‐CIPK26‐mediated phosphorylation enhances activity of the NADPH oxidase RBOHC, but is dispensable for root hair growth

Cited by 32 publications

References 49 publications

RBOH-Dependent ROS Synthesis and ROS Scavenging by Plant Specialized Metabolites To Modulate Plant Development and Stress Responses

RBOH-Dependent ROS Synthesis and ROS Scavenging by Plant Specialized Metabolites To Modulate Plant Development and Stress Responses

Fine‐tuning of RBOHF activity is achieved by differential phosphorylation and Ca²⁺ binding

Dual-sensing genetically encoded fluorescent indicators resolve the spatiotemporal coordination of cytosolic abscisic acid and second messenger dynamics in Arabidopsis

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CBL1‐CIPK26‐mediated phosphorylation enhances activity of the NADPH oxidase RBOHC, but is dispensable for root hair growth

Cited by 32 publications

References 49 publications

RBOH-Dependent ROS Synthesis and ROS Scavenging by Plant Specialized Metabolites To Modulate Plant Development and Stress Responses

RBOH-Dependent ROS Synthesis and ROS Scavenging by Plant Specialized Metabolites To Modulate Plant Development and Stress Responses

Fine‐tuning of RBOHF activity is achieved by differential phosphorylation and Ca2+ binding

Dual-sensing genetically encoded fluorescent indicators resolve the spatiotemporal coordination of cytosolic abscisic acid and second messenger dynamics in Arabidopsis

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Fine‐tuning of RBOHF activity is achieved by differential phosphorylation and Ca²⁺ binding