ABSTRACT: Arabidopsis floral initiator SKB1 confers high salt tolerance by regulating transcription and pre-mRNA splicing through altering histone H4R3 and small nuclear ribonucleoprotein LSM4 methylation
Plants adapt their growth and development in response to perceived salt stress. Although DELLA-dependent growth restraint is thought to be an integration of the plant's response to salt stress, little is known about how histone modification confers salt stress and, in turn, affects development. Here, we report that floral initiator Shk1 kinase binding protein1 (SKB1) and histone4 arginine3 (H4R3) symmetric dimethylation (H4R3sme2) integrate responses to plant developmental progress and salt stre ...[more]
Project description:This study examines how histone modification confers salt stress in Arabidopsis. The floral initiator SKB1 is found to mediate the plant’s response to salt stress by altering the methylation status of histone H4R3 and of the small nuclear ribonucleoprotein (snRNP) LSM4, and thereby affecting the expression of stress-responsive genes. Overall design: Total RNA was isolated with Trizol reagent (Invitrogen) from eleven11-day-old seedlings of the wild type and skb1-1 mutant without or with 200 mM NaCl for 6 hrs. 29k Arabidopsis Genome Array hybridization was carried out by CapitalBio Corporation (Beijing, China).
Project description:This study examines how histone modification confers salt stress in Arabidopsis. The floral initiator SKB1 is found to mediate the plant’s response to salt stress by altering the methylation status of histone H4R3 and of the small nuclear ribonucleoprotein (snRNP) LSM4, and thereby affecting the expression of stress-responsive genes. Total RNA was isolated with Trizol reagent (Invitrogen) from eleven11-day-old seedlings of the wild type and skb1-1 mutant without or with 200 mM NaCl for 6 hrs. 29k Arabidopsis Genome Array hybridization was carried out by CapitalBio Corporation (Beijing, China).
Project description:Generally, salt stress causes both osmotic and ionic stress. To discern the effects of osmotic and ionic specific effects on Burma mangrove transcriptome, we conducted expression profiling in 500 mM NaCl or 1M solbitol treated leaves. This study will lead to a rapid and effective selection of gene that confers high salt tolerance in transgenic plants and to a comprehensive understanding of plant stress response. Keywords: Stress response Overall design: The 19 hybridizations were carried out, 18 hybridizations in 6 conditions (NaCl treated for [0h, 6h, 1d], and Solbitol treated for [0h, 6h, 1d]) with 3 biological replicates [A,B,C] per condition) and 1 additional hybridization for the initial self- vs. self-hybridization to assess technical variability.
Project description:Loss of the seed-specific WRKY transcription factor WRKY43 confers enhanced tolerance towards high salt, high osmolarity and low temperature with respect to seed germination. wrky43 loss of function lines display increased inhibition of seed germination in response to exogenous ABA, while WRKY43 overexpression lines are more tolerant towards exogenous ABA. The opposing effect of the wrky43 mutant on salt and ABA tolerance is reminiscent of fatty acid desaturase mutants. Loss of WRKY43 enhances polyunsaturated fatty acid content, particularly 18:2 and 18:3 in TAGs and Phospholipids. Gene chip arrays show that ABA-induced regulation of FUSCA3, ZAT10 and seed storage proteins are absent in the wrky43 mutant. Promoter-Luciferase studies confirm direct regulation of ZAT10 by WRKY43 and suggest indirect regulation of FUS3 and SSPs. In summary WRKY43 acts as a positive regulator of ABA-dependent gene regulation and of fatty acid desaturation that finally results in enhanced tolerance to abiotic stress. 2 biological replicates of Arabidopsis thaliana Ler-0 wildtype and wrky43 muatnt seeds were compared after incubation in liquid 0.5 MS media with 2 µM ABA for 4 days
Project description:Biogenesis of eukaryotic messenger ribonucleoprotein complexes (mRNPs) involves the synthesis, splicing, and 3’-processing of pre-mRNA, and the assembly of mature mRNPs for nuclear export. We mapped 23 mRNP biogenesis factors onto the newly synthesized yeast transcriptome, providing ~10^5-10^6 high-confidence RNA interaction sites per factor. PAR-CLIP data of 23 mRNP biogenesis factors in Saccharomyces cerevisiae
Project description:Plant basic helix-loop-helix (bHLH) transcription factors are involved in physiological and developmental processes, and also play essential roles in abiotic stresses. However, their exact roles in abiotic stress are still need to be elucidated, and most of bHLHs have not been functionally characterized. In the present study, we characterized the functional role of AtbHLH112 in response to abiotic stresses. AtbHLH112 is a nuclear-localized protein, and its nuclear-localization is induced by salt, drought and ABA. Besides binding to E-box motif, AtbHLH112 is found to bind to a novel motif with the sequence “GG[GT]CC[GT][GA][TA]C” (GCG-box), and the binding affinity is induced by salt and ABA. Gain- and loss-of-function analyses showed that the transcript level of AtbHLH112 is positively correlated with salt and drought tolerance. AtbHLH112 mediates stress tolerance by upregulating the expression of P5CS genes and decreasing the expression of P5CDH and PRODH genes to increase proline levels, and via enhancing the expression of POD and SOD genes to improve ROS scavenging ability. All data together suggested that AtbHLH112 regulates the expression of genes through binding to GCG-box and E-box to mediate the physiological stress responses, including proline biosynthesis and ROS scavenging pathways to enhance stress tolerance. Differentially expression genes of AtbHLH112-overexpression plants, mutant (SALK_033618C) plants and wild type of Columbia Arabidopsis thaliana were measured under salt stressed and normal condition for 3 hours, respectively. Three independent experiments were performed at each treatment using different plants for each experiment.
Project description:We performed that comprehensive identification of genes responsible for stress tolerance by analyzing the whole-genome expression profiles of poplar (Populus alba × P. glandulosa) leaves exposed to drought and salt stresses. Examination at the molecular level how this tree species responds to drought and salt stresses by regulating the expression of genes involved in signal transduction, transcriptional regulation, and stress responses. Genome-wide analysis was conducted in poplar leaves exposed to drought and salt stresses.The plants were acclimated in soil and grown for 6 weeks in controlled conditions in a growth room (16 h light; light intensity, 150 μmol m-2sec-1; 24°C). Plants with a height of about 15 cm were separately exposed to either drought or salt stress. Up- and down-regulated genes were identified, and their putative functions are discussed.
Project description:Alternative pre-messenger RNA (pre-mRNA) splicing is a post-transcriptional mechanism for controlling gene expression. Splicing patterns are determined by both RNA binding proteins and nuclear pre-mRNA structure. Here, we analyze pre-mRNA splicing patterns, RNA binding sites and RNA structures near these binding sites coordinately controlled by two splicing factors, the heterogeneous nuclear ribonucleoprotein hnRNPA1 and the RNA helicase DDX5. We identified thousands of alternative pre-mRNA splicing events controlled by these factors by RNA-seq following RNA interference. Enhanced CLIP (eCLIP) on nuclear extracts was used to identify protein-RNA binding sites for both proteins in the nuclear transcriptome. We found a significant overlap between hnRNPA1 and DDX5 splicing targets and that they share many closely linked binding sites as determined by eCLIP analysis. In vivo SHAPE chemical RNA structure probing data was used to model RNA structures near several exons controlled and bound by both proteins. Both sequence motifs and in vivo UV crosslinking sites for hnRNPA1 and DDX5 were used to map binding sites in their RNA targets and often these sites flanked regions of higher chemical reactivity suggesting an organized nature to nuclear pre-mRNPs. This work provides a first glimpse into the possible RNA structures surrounding pre-mRNA splicing factor binding sites. Overall design: A human myeloid leukemia cell line, K562 cells, were treated with hnRNPA1 and DDX5 siRNA. Two replicate samples of hnRNPA1 knockdown and DDX5 knockdown were prepared for RNA-seq for differential pre-mRNA splicing analysis, along with control scramble siRNA samples. Triplicate eCLIP samples were prepared for hnRNPA1 and DDX5 using nuclear extracts in K562 cells, with input control and no UV treatment control samples.
Project description:Two groups of rats were fed either a high salt diet or a low salt diet. This study aims to look at salt intake in correlation to altering other metabolites and the onset of hypertension