Dataset Information

Sulfide Regulates Autophagy by Reversible Persulfidation of the Cysteine Protease ATG4

ABSTRACT: Hydrogen sulfide is a signaling molecule that regulates essential processes for plant performance, such as autophagy. In Arabidopsis thaliana, hydrogen sulfide negatively regulates autophagy independently of reactive oxygen species, but the underlying mechanism remains to be understood. To determine whether persulfidation, an emergent posttranslational modification, has a main role in the control of autophagy by sulfide, we have used a proteomic approach targeting ATG4, a cysteine protease that plays a crucial role in autophagy progression. We showed that AtATG4a from A. thaliana, which is the predominant ATG4 protease in this plant species, contains a specific site for persulfidation, the residue Cys170, included in the characteristic catalytic triad Cys-His-Asp of cysteine proteases. We tested whether persulfidation regulates the activity of ATG4 by setting up a heterologous assay using the Chlamydomonas reinhardtii CrATG8 protein as a substrate. Our findings demonstrate that sulfide significantly inactivates AtATG4a cleavage activity in a reversible manner. The biological significance of the reversible inhibition of the ATG4 protease by sulfide is supported by our findings in Arabidopsis leaves under both basal and autophagy-inducing conditions. We also observed a significant increase in the overall ATG4 activity in Arabidopsis under nitrogen starvation and osmotic stress, which is inhibited by sulfide. Therefore, our data strongly suggest that the negative regulation of autophagy by sulfide is mediated, at least, by the specific persulfidation of the protease ATG4.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Leaf

SUBMITTER: Luis C. Romero

LAB HEAD: Luis C Romero

PROVIDER: PXD019802 | Pride | 2020-12-14

REPOSITORIES: Pride

ACCESS DATA

Publications

Abscisic Acid-Triggered Persulfidation of the Cys Protease ATG4 Mediates Regulation of Autophagy by Sulfide.

Laureano-Marín Ana M AM Aroca Ángeles Á Pérez-Pérez M Esther ME Yruela Inmaculada I Jurado-Flores Ana A Moreno Inmaculada I Crespo José L JL Romero Luis C LC Gotor Cecilia C

The Plant cell 20201009 12

Hydrogen sulfide is a signaling molecule that regulates essential processes in plants, such as autophagy. In Arabidopsis (<i>Arabidopsis thaliana</i>), hydrogen sulfide negatively regulates autophagy independently of reactive oxygen species via an unknown mechanism. Comparative and quantitative proteomic analysis was used to detect abscisic acid-triggered persulfidation that reveals a main role in the control of autophagy mediated by the autophagy-related (ATG) Cys protease AtATG4a. This proteas ...[more]

PMID: 33037147

Similar Datasets

Project description:Hydrogen sulfide is an important signaling molecule in plants that regulates essential biological processes through protein persulfidation. Although numerous persulfidated proteins were found to be involved in photorespiration, little was known about sulfide-mediated regulation of photorespiration. Label-free quantitative proteomic analysis has revealed a high impact on the protein persulfidation level when plants grown under non-photorespiratory conditions are transferred to air, with the 97 % of total identified proteins more persulfidated under suppressed photorespiration. A detailed study of the effect of sulfide on important aspects associated with photorespiratory growth conditions has provided insight into the role of sulfide in protecting plants grown under suppressed photorespiration. In these conditions, sulfide amends the metabolic imbalance of carbon/nitrogen and restores ATP levels to concentrations similar to those in air grown conditions. Sulfide also plays an essential role in balancing the significant high level of ROS measured in plants under non-photorespiratory conditions to reach a similar cellular redox state in air-grown plants, through the regulation of antioxidative defenses. Furthermore, sulfide regulates another important characteristic resulting from suppression of photorespiration, such as the stomata closure, to decrease the high guard cell ROS levels and inducing the stomata aperture. In this way, sulfide signals the movement of the CO2-dependent stomata to achieve a scenario similar to that of plants growing under normal air conditions, in the opposite direction of the already established ABA-dependent movement of the stomata. Therefore, our findings suggest that the high persulfidation level under suppressed photorespiration reveals an essential role of sulfide signaling under these conditions, and with respect to stomatal movement, the outcome of this signaling is dependent on the growth/stress condition under study.

Project description:Hydrogen sulfide (H2S) is a signaling molecule that regulates essential plant processes, such as autophagy and responses to abiotic stresses. Although there is much information about the processes in which H2S is involved, the mechanism of action of H2S is still unclear. However, the posttranslational modification of cysteine residues, named persulfidation, is the main proposed mechanism. In this study, the role of H2S during drought that allows plant stress adaptation has been analyzed, with the focus on the underlying mechanism. H2S pretreatment before imposing drought on plants significantly improved phenotypic traits and decreased the content of biochemical and molecular drought stress markers. In addition, H2S regulated amino acid metabolism and repressed drought-induced degradation pathways, such as bulk autophagy and protein ubiquitination. The label-free quantitative proteomic analysis of plants growing under control and drought stress identified 887 proteins significantly different persulfidated between both conditions. Bioinformatic analyses revealed that the biological processes most enriched containing the proteins more persulfidated in drought corresponded to cellular response to oxidative stress, and hydrogen peroxide catabolism. In addition, protein degradation, abiotic stress responses, anthocyanin-containing compound biosynthetic process, and flavonoid biosynthesis were highlighted. On the contrary, the most enriched biological process containing proteins with lower levels of persulfidation in drought corresponded to the biosynthetic processes of glucosinolates and chlorophyll. Therefore, our findings emphasize the role of H2S as a promoter of enhanced tolerance to drought, enabling plants to respond more rapidly and efficiently after exposure to drought. Furthermore, the proteomic analysis supports the main role of protein persulfidation as the molecular mechanism of H2S to alleviate ROS accumulation and balance redox homeostasis under drought stress.

Dataset Information

Sulfide Regulates Autophagy by Reversible Persulfidation of the Cysteine Protease ATG4

Publications

Abscisic Acid-Triggered Persulfidation of the Cys Protease ATG4 Mediates Regulation of Autophagy by Sulfide.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure