Ethylene insensitive 2 (EIN2) destiny shaper: The post-translational modification.

PTMs (Post-Translational Modifications) of proteins facilitate rapid modulation of protein function in response to various environmental stimuli. The EIN2 (Ethylene Insensitive 2) protein is a core regulatory of the ethylene signaling pathway. Recent findings have demonstrated that PTMs, including protein phosphorylation, ubiquitination, and glycosylation, govern EIN2 trafficking, subcellular localization, stability, and physiological roles. The cognition of multiple PTMs in EIN2 underscores the stringent regulation of protein. Consequently, a thorough review of the regulatory role of PTMs in EIN2 functions will improve our profound comprehension of the regulation mechanism and various physiological processes of EIN2-mediated signaling pathways. This review discusses the evolution, functions, structure and characteristics of EIN2 protein in plants. Additionally, this review sheds light on the progress of protein ubiquitination, phosphorylation, O-Glycosylation in the regulation of EIN2 functions, and the unresolved questions and future perspectives.

Transcriptome analysis reveals the main metabolic pathway of c-GMP induced by salt stress in tomato (Solanum lycopersicum) seedlings.

Tomato (Solanum lycopersicum L.) is a model crop as well as an important food worldwide. In arid areas, increasing soil salinity has limited higher yields in tomato production. As a second messenger molecule, cyclic guanosine monophosphate (c-GMP) plays an indispensable role in plant response to salt stress by regulating cell processes to promote plant growth and development. However, this mechanism has not been fully explored in tomato seedlings. In this experiment, tomato seeds were cultured in four treatments: (1) distilled water (CK); (2) 20µM c-GMP (T1); (3) 50mM NaCl (T2); and (4) 20µM c-GMP+50mM NaCl (T3). The results show that 20µM c-GMP effectively alleviated the inhibitory effect of 50mM NaCl on growth and development, and induced the expression of 1580 differentially expressed genes (DEGs). Seedlings in the CK vs T1 shared 95 upregulated and 442 downregulated DEGs, whereas T2 vs T3 shared 271 upregulated and 772 downregulated DEGs. Based on KEGG (Kyoto Encyclopaedia of Genes and Genomes) analysis, the majority of DEGs were involved in metabolism; exogenous c-GMP induced significant enrichment of pathways associated with carbohydrates, phenylpropanoids and fatty acid metabolism. Most PMEs , acCoA , PAL , PODs , FADs , and AD were upregulated, and GAPDHs , PL , PG , BXL4 , and ß-G were downregulated, which reduced susceptibility of tomato seedlings to salt and promoted their salt tolerance. The application of c-GMP increased soluble sugar, flavonoid and lignin contents, reduced accumulation of malondialdehyde (MDA), and enhanced the activity of peroxidase (POD). Thus, our results provide insights into the molecular mechanisms associated with salt tolerance of tomato seedlings.

Type one protein phosphatase regulates fixed-carbon starvation-induced autophagy in Arabidopsis.

Autophagy, a conserved pathway that carries out the bulk degradation of cytoplasmic material in eukaryotic cells, is critical for plant physiology and development. This process is tightly regulated by ATG13, a core component of the ATG1 kinase complex, which initiates autophagy. Although ATG13 is known to be dephosphorylated immediately after nutrient starvation, the phosphatase regulating this process is poorly understood. Here, we determined that the Arabidopsis (Arabidopsis thaliana) septuple mutant (topp-7m) and octuple mutant (topp-8m) of TYPE ONE PROTEIN PHOSPHATASE (TOPP) exhibited significantly reduced tolerance to fixed-carbon (C) starvation due to compromised autophagy activity. Genetic analysis placed TOPP upstream of autophagy. Interestingly, ATG13a was found to be an interactor of TOPP. TOPP directly dephosphorylated ATG13a in vitro and in vivo. We identified 18 phosphorylation sites in ATG13a by LC-MS. Phospho-dead ATG13a at these 18 sites significantly promoted autophagy and increased the tolerance of the atg13ab mutant to fixed-C starvation. The dephosphorylation of ATG13a facilitated ATG1a-ATG13a complex formation. Consistently, the recruitment of ATG13a for ATG1a was markedly inhibited in topp-7m-1. Finally, TOPP-controlled dephosphorylation of ATG13a boosted ATG1a phosphorylation. Taken together, our study reveals the crucial role of TOPP in regulating autophagy by stimulating the formation of the ATG1a-ATG13a complex by dephosphorylating ATG13a in Arabidopsis.

Enzymatic synthesis of PEGylated lactide-diester-diol copolyesters for highly efficient targeted anticancer drug delivery.

PEGylated lactide-diester-diol copolymers were successfully synthesized via lipase-catalyzed copolymerization, the resultant amphiphilic PEG-poly(L-lactate-co-hexamethylene-co-adipate) (PEG-PLLHA) and PEG-poly(D,L-lactate-co-hexamethylene-co-adipate) (PEG-PDLLHA) block copolymers readily undergo self-assembly processes to form nanosized micelles in aqueous medium, which are stable under physiological conditions in the presence of serum proteins. By conjugating folic acid (FA) to enzymatic synthesized poly(hexamethylene adipate-co-hexamethylene 2,3-epoxy succinate), we could formulate FA-bearing PEG-polyester micelles for docetaxel (DTX) targeting delivery. FA-PEG-PLLHA and FA-PEG-PDLLHA micelles possess efficient cell-targeting capability toward FA receptor-positive cancer cells (e.g., CT-26), which significantly enhances their cellular uptake rates and efficacy of drug-loaded formulations toward such cells. During in vivo anticancer treatments, the FA-bearing micelles are highly capable of targeting and accumulating preferentially in tumor tissues by both active cell-targeting mechanism and passive targeting via the EPR effect. All these desirable properties enable the FA-bearing micelles to deliver DTX with 97% tumor-inhibiting efficiency through systemic delivery, which is favorable in comparison to the values of various DTX nanoparticle formulations reported in literature. Importantly, biosafety assays reveal that all DTX-loaded micelles are biocompatible and safe for in vivo antitumor treatment applications. Thus, FA-PEG-PLLHA and FA-PEG-PDLLHA micelles represent new types of promising anticancer drug nanocarriers for targeted chemotherapy.

Enzymatic multifunctional biodegradable polymers for pH- and ROS-responsive anticancer drug delivery.

A new family of multifunctional biodegradable block copolymers, PEG-poly(ω-pentadecalactone-co-N-methyldiethyleneamine sebacate-co-2,2'-thiodiethylene sebacate) (PEG-PMT), were synthesized via lipase-catalyzed copolymerization procedures. Amphiphilic PEG-PMT copolymers can be readily transformed into stable micellar nanoparticles through self-assembling processes in aqueous medium. The particle sizes increase dramatically after exposure of the particles to the acidic pH and high reactive oxygen species (ROS) conditions in tumor microenvironments, due to protonation of thioether groups and oxidation of amino groups in the PMT micelle cores, respectively. For example, docetaxel (DTX)-loaded PEG-PM-19 % TS micelles were triggered synergistically by acidic pH and ROS stimuli to release over 85 % of the anti-cancer drug. In particular, DTX/PEG-PMT-19 % TS and DTX/PEG-PMT-48 % TS micelles performed better than commercial Duopafei formulation in prohibiting growth of CT-26 tumors xenografed in vivo (70 % of tumor-inhibiting efficiency). Biosafety analysis revealed that DTX-loaded PEG-PMT nanoparticles possessed minimal toxicity towards normal organs, such as liver and kidney. These experimental data demonstrated that the pH- and ROS-responsive PEG-PMT micelles are promising vectors for both delivery of anti-tumor drugs and their controlled release at tumor intracellular sites.

Identification of Exogenous Nitric Oxide-Responsive miRNAs from Alfalfa (Medicago sativa L.) under Drought Stress by High-Throughput Sequencing.

Alfalfa (Medicago sativa L.) is a high quality leguminous forage. Drought stress is one of the main factors that restrict the development of the alfalfa industry. High-throughput sequencing was used to analyze the microRNA (miRNA) profiles of alfalfa plants treated with CK (normal water), PEG (polyethylene glycol-6000; drought stress), and PEG + SNP (sodium nitroprusside; nitric oxide (NO) sprayed externally under drought stress). We identified 90 known miRNAs belonging to 46 families and predicted 177 new miRNAs. Real-time quantitative fluorescent PCR (qRT-PCR) was used to validate high-throughput expression analysis data. A total of 32 (14 known miRNAs and 18 new miRNAs) and 55 (24 known miRNAs and 31 new miRNAs) differentially expressed miRNAs were identified in PEG and PEG + SNP samples. This suggested that exogenous NO can induce more new miRNAs. The differentially expressed miRNA maturation sequences in the two treatment groups were targeted by 86 and 157 potential target genes, separately. The function of target genes was annotated by gene ontology (GO) enrichment and kyoto encyclopedia of genes and genomes (KEGG) analysis. The expression profiles of nine selected miRNAs and their target genes verified that their expression patterns were opposite. This study has documented that analysis of miRNA under PEG and PEG + SNP conditions provides important insights into the improvement of drought resistance of alfalfa by exogenous NO at the molecular level. This has important scientific value and practical significance for the improvement of plant drought resistance by exogenous NO.

Enzymatic PEG-Poly(amine-co-disulfide ester) Nanoparticles as pH- and Redox-Responsive Drug Nanocarriers for Efficient Antitumor Treatment.

We have designed and constructed novel multifunctional nanoparticle drug-delivery systems that are stable under physiological conditions and responsive to tumor-relevant pH and intracellular reduction potential. The nanoparticles were fabricated from enzymatically synthesized poly(ethylene glycol) (PEG)-poly(ω-pentadecalactone-co-N-methyldiethyleneamine-co-3,3'-dithiodipropionate) (PEG-PPMD) and PEG-poly(ε-caprolactone-co-N-methyldiethyleneamine-co-3,3'-dithiodipropionate) (PEG-PCMD) block copolymers via self-assembly processes in aqueous solution. At acidic pH and in the presence of a reductant (e.g., d,l-dithiothreitol or glutathione), the nanosized micelle particles rapidly swell and disintegrate due to the protonation of amino groups and reductive cleavage of disulfide bonds in the micelle cores. Consistently, docetaxel (DTX)-loaded PEG-PPMD and PEG-PCMD micelles can be triggered synergistically by acidic endosomal pH and a high intracellular reduction potential to rapidly release the drug for efficient killing of cancer cells. The drug formulations based on PEG-PPMD and PEG-PCMD copolymers exhibited a substantially higher potency than free DTX in inhibiting tumor growth in mice, whereas their therapeutic effects on important organ tissues were minimal. These results demonstrate that PEG-PPMD and PEG-PCMD nanoparticles have a great potential to serve as site-specific, controlled drug-delivery vehicles for safe and efficient antitumor treatment.