Short-Term Low Temperature Induces Nitro-Oxidative Stress that Deregulates the NADP-Malic Enzyme Function by Tyrosine Nitration in Arabidopsis thaliana

Abstract

Low temperature (LT) negatively affects plant growth and development via the alteration of the metabolism of reactive oxygen and nitrogen species (ROS and RNS). Among RNS, tyrosine nitration, the addition of an NO2 group to a tyrosine residue, can modulate reduced nicotinamide-dinucleotide phosphate (NADPH)-generating systems and, therefore, can alter the levels of NADPH, a key cofactor in cellular redox homeostasis. NADPH also acts as an indispensable electron donor within a wide range of enzymatic reactions, biosynthetic pathways, and detoxification processes, which could affect plant viability. To extend our knowledge about the regulation of this key cofactor by this nitric oxide (NO)-related post-translational modification, we analyzed the effect of tyrosine nitration on another NADPH-generating enzyme, the NADP-malic enzyme (NADP-ME), under LT stress. In Arabidopsis thaliana seedlings exposed to short-term LT (4 °C for 48 h), a 50% growth reduction accompanied by an increase in the content of superoxide, nitric oxide, and peroxynitrite, in addition to diminished cytosolic NADP-ME activity, were found. In vitro assays confirmed that peroxynitrite inhibits cytosolic NADP-ME2 activity due to tyrosine nitration. The mass spectrometric analysis of nitrated NADP-ME2 enabled us to determine that Tyr-73 was exclusively nitrated to 3-nitrotyrosine by peroxynitrite. The in silico analysis of the Arabidopsis NADP-ME2 protein sequence suggests that Tyr73 nitration could disrupt the interactions between the specific amino acids responsible for protein structure stability. In conclusion, the present data show that short-term LT stress affects the metabolism of ROS and RNS, which appears to negatively modulate the activity of cytosolic NADP-ME through the tyrosine nitration process.

Oxidative Stress in Plants

“…These authors suggested that inoculation with the Serratia marcescens BM1 would promotes Cd stress tolerance and phytoremediation potential. The impact of abiotic stress was also reported by Dr. Barroso’s group [ 2 ] as they demonstrated the effect of short-term low temperature stress on the metabolism of reactive oxygen and nitrogen species in Arabidopsis plants. These authors showed that the low temperature produces nitro-oxidative stress, and reduces cytosolic NADP-malic enzyme activity, which was negatively modulated by the protein tyrosine nitration process.…”

“…(1) El-Esawi et al, 2020 [ 1 ]. (2) Begara-Morales et al, 2019 [ 2 ]. (3) García-Martí et al, 2019 [ 3 ].…”

Assessment of Subcellular ROS and NO Metabolism in Higher Plants: Multifunctional Signaling Molecules

“…NADPH-generating enzymes maintain redox state and support antioxidant enzymes, such as GR, as well as O 2 •− and NO generation by RBOH and NOS-like activity, respectively. These NADP-DHs are differentially modulated by nitration/S-nitrosation: chloroplast FNR is inhibited by nitration, NADP-ICDH and NADP-ME are inhibited by both nitration and S-nitrosation [173], while G6PDH and 6PGDH appear unaffected [174].…”

Tomato leaves under stress: A comparison of stress response to mild abiotic stress between a cultivated and a wild tomato species

“…Many environmental stresses, result in oxidative stress (Abewoy Fentik, 2017;Ksas et al, 2015;Chokshi et al, 2017;Robles-Rengel et al, 2019;Munjal and Munjal, 2019;Begara-Morales et al, 2019;García-Martí et al, 2019). As an often shown consequence, biosynthesis of plant secondary metabolites (PSM) is stimulated (Løvdal et al, 2010;Selmar and Kleinwächter, 2013;Grace, 2007;Dixon and Paiva, 1995) to overcome the stress (Wu et al, 2014;Isah, 2019;Aguirre-Becerra et al, 2021) and / or to become more tolerant (Nakabayashi et al, 2014).…”