Proteome-wide and lysine crotonylation profiling reveals the importance of crotonylation in chrysanthemum (Dendranthema grandiforum) under low-temperature

Abstract

Background Low-temperature severely affects the growth and development of chrysanthemum which is one kind of ornamental plant well-known and widely used in the world. Lysine crotonylation is a recently identified post-translational modification (PTM) with multiple cellular functions. However, lysine crotonylation under low-temperature stress has not been studied. Results Proteome-wide and lysine crotonylation of chrysanthemum at low-temperature was analyzed using TMT (Tandem Mass Tag) labeling, sensitive immuno-precipitation, and high-resolution LC-MS/MS. The results showed that 2017 crotonylation sites were identified in 1199 proteins. Treatment at 4 °C for 24 h and − 4 °C for 4 h resulted in 393 upregulated proteins and 500 downregulated proteins (1.2-fold threshold and P < 0.05). Analysis of biological information showed that lysine crotonylation was involved in photosynthesis, ribosomes, and antioxidant systems. The crotonylated proteins and motifs in chrysanthemum were compared with other plants to obtain orthologous proteins and conserved motifs. To further understand how lysine crotonylation at K136 affected APX (ascorbate peroxidase), we performed a site-directed mutation at K136 in APX. Site-directed crotonylation showed that lysine decrotonylation at K136 reduced APX activity, and lysine complete crotonylation at K136 increased APX activity. Conclusion In summary, our study comparatively analyzed proteome-wide and crotonylation in chrysanthemum under low-temperature stress and provided insights into the mechanisms of crotonylation in positively regulated APX activity to reduce the oxidative damage caused by low-temperature stress. These data provided an important basis for studying crotonylation to regulate antioxidant enzyme activity in response to low-temperature stress and a new research ideas for chilling-tolerance and freezing-tolerance chrysanthemum molecular breeding.

Atomically Dispersed Cu Nanozyme with Intensive Ascorbate Peroxidase Mimic Activity Capable of Alleviating ROS‐Mediated Oxidation Damage

“…[ 3 , 4 ] Though APX functions as the vital regulator for maintaining the ROS balance in plant cells, APX is susceptible to multiple redox‐related post‐transitional modifications (PTMs) and easily inactivated by specific inhibitors through blocking the oxidation of H 2 O 2 with heme. [ 4 , 5 ] Recently much effort has been put into regulating the activities of APXs in terms of post‐transitional modifications of the proteins, [ 6 , 7 ] it only succeeded marginally. Until now, for characterizing the regulation by PTMs, only a limited number of proteins have been investigated.…”

Qualitative Proteome-Wide Analysis Reveals the Diverse Functions of Lysine Crotonylation in Dendrobium huoshanense

“…However, there was evidence showing that crotonylation possesses a certain association to plant growth and development. With the innovation of high-resolution MS and highly specific antibodies, research on plant crotonylation has been greatly promoted, such as in peanut ( Xu et al, 2021 ), papaya ( Liu et al, 2018a ), Chrysanthemum ( Lin et al, 2021 ), tea ( Sun et al, 2019 ), rice ( Liu et al, 2018b ), and tobacco ( Sun et al, 2017 ; Table 1 ). However, there were no reports for crotonylation in D. huoshanense .…”

“…Based on conserved protein analysis ( Figures 6A–D ), APX, CAT, GPX, GRP2, and GST were conserved in tea, Chrysanthemum , rice, and tobacco, implying that all the five enzymes were extensively crotonylated in plants. Crotonylation had been shown to positively regulate APX activity under abiotic stress, further reduce the oxidative damage caused by it Lin et al (2021) . Therefore, the crotonylation of proteins may modulate ROS stress.…”

“…PTM is a more rapid and effective strategy than gene transcription and translation ( He et al, 2016 ). However, Kcr in plants had only been analyzed in rice ( Liu et al, 2018b ), tobacco ( Sun et al, 2017 ), papaya ( Liu et al, 2018a ), tea plants ( Sun et al, 2019 ), Chrysanthemum ( Lin et al, 2021 ), and peanut ( Xu et al, 2021 ) and was found to contribute to a range of processes, including photosynthesis, glycolysis, and amino acid synthesis.…”

Global crotonylatome and GWAS revealed a TaSRT1 - TaPGK model regulating wheat cold tolerance through mediating pyruvate

“…In rice, mitogen-activated protein kinase 3 (OsMAPK3) phosphorylates OsICE1 and inhibits its ubiquitination to activate trehalose-6-phosphate phosphatase (OsTPP1) and enhances chilling tolerance ( 15 ). As a novel lysine PTM, crotonylation (Kcr) was also found to be related to cold stress in plants ( 16 ). In chrysanthemum, Kcr of a temperature-induced lipocalin-1-like gene (DgTIL1) promoted the cold tolerance by inhibiting a nonspecific lipid transfer protein (DgnsLTP) degradation ( 17 ).…”