Muscle-specific Cand2 is translationally upregulated by mTORC1 and promotes adverse cardiac remodeling.

The mechanistic target of rapamycin (mTOR) promotes pathological remodeling in the heart by activating ribosomal biogenesis and mRNA translation. Inhibition of mTOR in cardiomyocytes is protective; however, a detailed role of mTOR in translational regulation of specific mRNA networks in the diseased heart is unknown. We performed cardiomyocyte genome-wide sequencing to define mTOR-dependent gene expression control at the level of mRNA translation. We identify the muscle-specific protein Cullin-associated NEDD8-dissociated protein 2 (Cand2) as a translationally upregulated gene, dependent on the activity of mTOR. Deletion of Cand2 protects the myocardium against pathological remodeling. Mechanistically, we show that Cand2 links mTOR signaling to pathological cell growth by increasing Grk5 protein expression. Our data suggest that cell-type-specific targeting of mTOR might have therapeutic value against pathological cardiac remodeling.

Identification of novel proteins for lacunar stroke by integrating genome-wide association data and human brain proteomes.

BACKGROUND: Previous genome-wide association studies (GWAS) have identified numerous risk genes for lacunar stroke, but it is challenging to decipher how they confer risk for the disease. We employed an integrative analytical pipeline to efficiently transform genetic associations to identify novel proteins for lacunar stroke. METHODS: We systematically integrated lacunar stroke genome-wide association study (GWAS) (N=7338) with human brain proteomes (N=376) to perform proteome-wide association studies (PWAS), Mendelian randomization (MR), and Bayesian colocalization. We also used an independent human brain proteomic dataset (N=152) to annotate the new genes. RESULTS: We found that the protein abundance of seven genes (ICA1L, CAND2, ALDH2, MADD, MRVI1, CSPG4, and PTPN11) in the brain was associated with lacunar stroke. These seven genes were mainly expressed on the surface of glutamatergic neurons, GABAergic neurons, and astrocytes. Three genes (ICA1L, CAND2, ALDH2) were causal in lacunar stroke (P < 0.05/proteins identified for PWAS; posterior probability of hypothesis 4 ≥ 75 % for Bayesian colocalization), and they were linked with lacunar stroke in confirmatory PWAS and independent MR. We also found that ICA1L is related to lacunar stroke at the brain transcriptome level. CONCLUSIONS: Our present proteomic findings have identified ICA1L, CAND2, and ALDH2 as compelling genes that may give key hints for future functional research and possible therapeutic targets for lacunar stroke.

CAND2/PMTR1 Is Required for Melatonin-Conferred Osmotic Stress Tolerance in Arabidopsis.

Osmotic stress severely inhibits plant growth and development, causing huge loss of crop quality and quantity worldwide. Melatonin is an important signaling molecule that generally confers plant increased tolerance to various environmental stresses, however, whether and how melatonin participates in plant osmotic stress response remain elusive. Here, we report that melatonin enhances plant osmotic stress tolerance through increasing ROS-scavenging ability, and melatonin receptor CAND2 plays a key role in melatonin-mediated plant response to osmotic stress. Upon osmotic stress treatment, the expression of melatonin biosynthetic genes including SNAT1, COMT1, and ASMT1 and the accumulation of melatonin are increased in the wild-type plants. The snat1 mutant is defective in osmotic stress-induced melatonin accumulation and thus sensitive to osmotic stress, while exogenous melatonin enhances the tolerance of the wild-type plant and rescues the sensitivity of the snat1 mutant to osmotic stress by upregulating the expression and activity of catalase and superoxide dismutase to repress H2O2 accumulation. Further study showed that the melatonin receptor mutant cand2 exhibits reduced osmotic stress tolerance with increased ROS accumulation, but exogenous melatonin cannot revert its osmotic stress phenotype. Together, our study reveals that CADN2 functions necessarily in melatonin-conferred osmotic stress tolerance by activating ROS-scavenging ability in Arabidopsis.

Melatonin in business with abiotic stresses in plants.

Melatonin (MEL) is the potential biostimulator molecule, governing multiple range of growth and developmental processes in plants, particularly under different environmental constrains. Mainly, its role is considered as an antioxidant molecule that copes with oxidative stress through scavenging of reactive oxygen species and modulation of stress related genes. It also enhances the antioxidant enzyme activities and thus helps in regulating the redox hemostasis in plants. Apart from its broad range of antioxidant functions, it is involved in the regulation of various physiological processes such as germination, lateral root growth and senescence in plants. Moreover this multifunctional molecule takes much interest due to its recent identification and characterization of receptorCandidate G-protein-Coupled Receptor 2/Phytomelatonin receptor(CAND2/PMTR1) in Arabidopsis thaliana. In this compiled work, different aspects of melatonin in plants such as melatonin biosynthesis and detection in plants, signaling pathway, modulation of stress related genes and physiological role of melatonin under different environmental stresses have been dissected in detail.

Phytomelatonin receptor PMTR1-mediated signaling regulates stomatal closure in Arabidopsis thaliana.

Melatonin has been detected in plants in 1995; however, the function and signaling pathway of this putative phytohormone are largely undetermined due to a lack of knowledge about its receptor. Here, we discovered the first phytomelatonin receptor (CAND2/PMTR1) in Arabidopsis thaliana and found that melatonin governs the receptor-dependent stomatal closure. The application of melatonin induced stomatal closure through the heterotrimeric G protein α subunit-regulated H2 O2 and Ca2+ signals. The Arabidopsis mutant lines lacking AtCand2 that encodes a candidate G protein-coupled receptor were insensitive to melatonin-induced stomatal closure. Accordingly, the melatonin-induced H2 O2 production and Ca2+ influx were completely abolished in cand2. CAND2 is a membrane protein that interacts with GPA1 and the expression of AtCand2 was tightly regulated by melatonin in various organs and guard cells. CAND2 showed saturable and specific 125 I-melatonin binding, with apparent Kd (dissociation constant) of 0.73 ± 0.10 nmol/L (r2  = .99), demonstrating this protein is a phytomelatonin receptor (PMTR1). Our results suggest that the phytomelatonin regulation of stomatal closure is dependent on its receptor CAND2/PMTR1-mediated H2 O2 and Ca2+ signaling transduction cascade.

Function, Mechanism, and Application of Plant Melatonin: An Update with a Focus on the Cereal Crop, Barley (Hordeum vulgare L.).

Melatonin is a multiple-function molecule that was first identified in animals and later in plants. Plant melatonin regulates versatile processes involved in plant growth and development, including seed germination, root architecture, flowering time, leaf senescence, fruit ripening, and biomass production. Published reviews on plant melatonin have been focused on two model plants: (1) Arabidopsis and (2) rice, in which the natural melatonin contents are quite low. Efforts to integrate the function and the mechanism of plant melatonin and to determine how plant melatonin benefits human health are also lacking. Barley is a unique cereal crop used for food, feed, and malt. In this study, a bioinformatics analysis to identify the genes required for barley melatonin biosynthesis was first performed, after which the effects of exogenous melatonin on barley growth and development were reviewed. Three integrated mechanisms of melatonin on plant cells were found: (1) serving as an antioxidant, (2) modulating plant hormone crosstalk, and (3) signaling through a putative plant melatonin receptor. Reliable approaches for characterizing the function of barley melatonin biosynthetic genes and to modulate the melatonin contents in barley grains are discussed. The present paper should be helpful for the improvement of barley production under hostile environments and for the reduction of pesticide and fungicide usage in barley cultivation. This study is also beneficial for the enhancement of the nutritional values and healthcare functions of barley in the food industry.

Analysis of DNA Methylation and Hydroxymethylation in the Genome of Crustacean Daphnia pulex.

The aim of our study was to analyze the presence of 5-methyl-cytosine (5-mC) and 5-hydroxymethyl-cytosine (5-hmC) in the genome of crustacean Daphnia pulex. First, the presence of 5-mC and 5-hmC in genomic DNA was demonstrated by using antibodies specific to either 5-mC or 5-hmC. Then, analysis of 5-mC and 5-hmC using pairs of restriction enzymes with different sensitivity to methylation and hydroxymethylation confirmed the presence of both modifications in selected regions of three genes (Cox4, Cand2 and Ephx1). To get a detailed picture of 5-hmC distribution over the D. pulex genome, we performed 5-hmC enrichment and sequenced the enriched fraction using next generation sequencing and non-enriched library (input) as a control. Comparison of input and enriched libraries showed that 5-hmC in exons is twice as frequent as in introns. Functional analysis indicated that 5-hmC abundance is associated with genes that are involved in the adenylate cyclase-activating G-protein-coupled receptor signaling pathway, molting cycles, morphogenesis and cell fate determination. Genes that lack 5-hmC tend to be involved in the regulation of the transforming growth factor beta receptor signaling pathway and in many mRNA-related processes. Our results suggest that epigenetic modifications are present in the genome of D. pulex and most likely are involved in the regulation of gene expression of this crustacean.