transcriptome and proteome

转录组和蛋白质组
  • 文章类型: Journal Article
    细菌核糖核酸酶E(RNaseE)通过降解和加工RNA对转录后调控至关重要。RraA蛋白通过蛋白质-蛋白质相互作用抑制RNaseE活性,对基因表达发挥整体调节作用。然而,RraA的具体作用尚不清楚.在这项研究中,我们研究了rraA在溶藻弧菌ZJ-T中的表达,并鉴定了三个负责其表达的启动子,产生具有不同5'-UTR长度的转录本。在静止阶段,rraA在转录后被显著抑制。rraA的缺失对含盐的丰富培养基Luria-Bertani肉汤(LBS)中的细菌生长没有影响,但导致生物膜形成减少和对多粘菌素B的抗性增加。转录组分析显示,野生型和rraA突变体之间有350个差异表达基因(DEG),而蛋白质组分析鉴定出267种差异表达蛋白(DEP)。整合分析确定了DEGs和DEP共有的55个基因,这表明RraA主要在转录后水平影响基因表达。KEGG(京都基因和基因组百科全书)分析表明,RraA促进脂肪酸的转化,丙酸,和支链氨基酸乙酰辅酶A,同时增强氨基酸和肽的摄取。值得注意的是,RraA正调控毒力相关基因的表达,包括参与生物膜形成和VI型分泌系统的那些。本研究通过转录组分析扩展了对RraA调控网络的理解,强调蛋白质组学分析在研究转录后调控中的重要性。IMPORTANCERraA是核糖核酸酶E的抑制剂蛋白,与核酸内切酶相互作用并抑制其核酸内切活性,从而在多种mRNA和非编码小RNA的降解和成熟中起着广泛的调节作用。然而,RraA在溶藻弧菌中的生理功能和相关调节子尚未完全阐明。这里,我们报道RraA影响毒力相关的生理过程,即,抗生素耐药性和生物膜形成,在溶藻中。通过对转录组和蛋白质组进行综合分析,我们揭示了RraA参与碳代谢,氨基酸分解代谢,和运输,以及VI型分泌系统。总的来说,这些发现阐明了RraA对与溶藻弧菌代谢和发病机制相关的多种途径的调节作用.
    Bacterial ribonuclease E (RNase E) is vital for posttranscriptional regulation by degrading and processing RNA. The RraA protein inhibits RNase E activity through protein-protein interactions, exerting a global regulatory effect on gene expression. However, the specific role of RraA remains unclear. In this study, we investigated rraA expression in Vibrio alginolyticus ZJ-T and identified three promoters responsible for its expression, resulting in transcripts with varying 5\'-UTR lengths. During the stationary phase, rraA was significantly posttranscriptionally inhibited. Deletion of rraA had no impact on bacterial growth in rich medium Luria-Bertani broth with salt (LBS) but resulted in decreased biofilm formation and increased resistance to polymyxin B. Transcriptome analysis revealed 350 differentially expressed genes (DEGs) between the wild type and the rraA mutant, while proteome analysis identified 267 differentially expressed proteins (DEPs). Integrative analysis identified 55 genes common to both DEGs and DEPs, suggesting that RraA primarily affects gene expression at the posttranscriptional level. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis demonstrated that RraA facilitates the conversion of fatty acids, propionic acid, and branched-chain amino acids to acetyl-CoA while enhancing amino acid and peptide uptake. Notably, RraA positively regulates the expression of virulence-associated genes, including those involved in biofilm formation and the type VI secretion system. This study expands the understanding of the regulatory network of RraA through transcriptome analysis, emphasizing the importance of proteomic analysis in investigating posttranscriptional regulation.IMPORTANCERraA is an inhibitor protein of ribonuclease E that interacts with and suppresses its endonucleolytic activity, thereby playing a widespread regulatory role in the degradation and maturation of diverse mRNAs and noncoding small RNAs. However, the physiological functions and associated regulon of RraA in Vibrio alginolyticus have not been fully elucidated. Here, we report that RraA impacts virulence-associated physiological processes, namely, antibiotic resistance and biofilm formation, in V. alginolyticus. By conducting an integrative analysis of both the transcriptome and proteome, we revealed the involvement of RraA in carbon metabolism, amino acid catabolism, and transport, as well as in the type VI secretion system. Collectively, these findings elucidate the regulatory influence of RraA on multiple pathways associated with metabolism and pathogenesis in V. alginolyticus.
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  • 文章类型: Journal Article
    杂交水稻(Oryzasativa)在产量和抗逆性方面通常优于其近交亲本,一种被称为杂种优势的现象,但是潜在的机制还没有完全理解。这里,我们结合了转录组,蛋白质组,生理,和杂种优势分析,以检查超级杂交稻超优1000(CY1000)的盐响应。除了超过其两个亲本的平均值(中亲本杂种优势)之外,CY1000表现出比其两个亲本更高的活性氧清除能力(过亲本杂种优势或杂种优势)。非加性表达和等位基因特异性基因表达实验表明,谷胱甘肽S-转移酶基因OsGSTU26和氨基酸转运体基因OsAAT30可能在耐盐性杂种优势中起主要作用。在CY1000中以占主导地位的方式行事。此外,我们将OsWRKY72确定为一种常见的转录因子,可结合并调节OsGSTU26和OsAAT30.在核心水稻种质品种中,盐敏感性表型与OsWRKY72父系基因型或OsAAT30母系基因型相关。OsWRKY72paternal特异性抑制盐胁迫下OsGSTU26的表达,导致盐度敏感性,而OsWRKY72母体特异性抑制OsAAT30,导致耐盐性。这些结果表明,OsWRKY72-OsAAT30/OsGSTU26模块可能在CY1000杂交水稻的耐盐性杂种优势中发挥重要作用,为阐明杂交水稻耐盐性杂种优势的机理提供了有价值的线索。
    Hybrid rice (Oryza sativa) generally outperforms its inbred parents in yield and stress tolerance, a phenomenon termed heterosis, but the underlying mechanism is not completely understood. Here, we combined transcriptome, proteome, physiological, and heterosis analyses to examine the salt response of super hybrid rice Chaoyou1000 (CY1000). In addition to surpassing the mean values for its two parents (mid-parent heterosis), CY1000 exhibited a higher reactive oxygen species scavenging ability than both its parents (over-parent heterosis or heterobeltiosis). Nonadditive expression and allele-specific gene expression assays showed that the glutathione S-transferase gene OsGSTU26 and the amino acid transporter gene OsAAT30 may have major roles in heterosis for salt tolerance, acting in an overdominant fashion in CY1000. Furthermore, we identified OsWRKY72 as a common transcription factor that binds and regulates OsGSTU26 and OsAAT30. The salt-sensitive phenotypes were associated with the OsWRKY72paternal genotype or the OsAAT30maternal genotype in core rice germplasm varieties. OsWRKY72paternal specifically repressed the expression of OsGSTU26 under salt stress, leading to salinity sensitivity, while OsWRKY72maternal specifically repressed OsAAT30, resulting in salinity tolerance. These results suggest that the OsWRKY72-OsAAT30/OsGSTU26 module may play an important role in heterosis for salt tolerance in an overdominant fashion in CY1000 hybrid rice, providing valuable clues to elucidate the mechanism of heterosis for salinity tolerance in hybrid rice.
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  • 文章类型: Journal Article
    背景:ZosteramarinaL.,或者鳗草,是整个北半球温带最广泛的海草物种。与陆地植物的干燥种子不同,鳗草种子必须在水中生存,盐度是影响鳗草种子萌发的关键因素。在本研究中,结合转录组和蛋白质组分析,研究低盐度刺激鳗草种子萌发的机制,除了发芽下关键代谢途径的动力学。
    结果:根据结果,低盐度刺激Ca2+信号和磷脂酰肌醇信号的激活,并通过MAPK转导级联进一步启动了各种与发芽相关的生理过程。淀粉,脂质,和贮藏蛋白被积极动员,为发芽提供能量和物质基础;脱落酸的合成和信号转导被抑制,赤霉素的合成和信号转导被激活,削弱种子休眠,准备发芽;激活细胞壁弱化和重塑过程,为子叶突出提供保护;此外,多个抗氧化系统被激活以减轻发芽过程中产生的氧化应激;ERF转录因子在两个阶段中的数量最高,表明在鳗草种子萌发中具有积极作用。
    结论:总之,第一次,本研究探讨了低盐度刺激鳗草种子萌发的机制,并全面分析了鳗草种子萌发过程中的转录组学和蛋白质组学特征。本研究的结果增强了我们对海草种子萌发的理解,特别是海草种子的分子生态学。
    BACKGROUND: Zostera marina L., or eelgrass, is the most widespread seagrass species throughout the temperate northern hemisphere. Unlike the dry seeds of terrestrial plants, eelgrass seeds must survive in water, and salinity is the key factor influencing eelgrass seed germination. In the present study, transcriptome and proteome analysis were combined to investigate the mechanisms via which eelgrass seed germination was stimulated by low salinity, in addition to the dynamics of key metabolic pathways under germination.
    RESULTS: According to the results, low salinity stimulated the activation of Ca2+ signaling and phosphatidylinositol signaling, and further initiated various germination-related physiological processes through the MAPK transduction cascade. Starch, lipids, and storage proteins were mobilized actively to provide the energy and material basis for germination; abscisic acid synthesis and signal transduction were inhibited whereas gibberellin synthesis and signal transduction were activated, weakening seed dormancy and preparing for germination; cell wall weakening and remodeling processes were activated to provide protection for cotyledon protrusion; in addition, multiple antioxidant systems were activated to alleviate oxidative stress generated during the germination process; ERF transcription factor has the highest number in both stages suggested an active role in eelgrass seed germination.
    CONCLUSIONS: In summary, for the first time, the present study investigated the mechanisms by which eelgrass seed germination was stimulated by low salinity and analyzed the transcriptomic and proteomic features during eelgrass seed germination comprehensively. The results of the present study enhanced our understanding of seagrass seed germination, especially the molecular ecology of seagrass seeds.
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  • 文章类型: Journal Article
    洪水是严重影响植物生存和发展的主要环境压力之一。然而,洪水胁迫的调节机制在Myricarialaxiflora中仍然未知,主要分布在长江洪泛区的濒危植物,中国。在这项工作中,在洪水和洪水后恢复处理下的9个时间点中,在M.laxiflora的根中并行进行了转录组和蛋白质组。总的来说,在洪水和洪水后恢复处理中观察到基因/蛋白质的高度动态和阶段特异性表达谱。与生长素相关的基因,细胞壁,钙信号,在洪水的早期阶段,MAP激酶信号仅在转录组水平上被大大下调。糖酵解和主要的CHO代谢基因,在转录组和/或蛋白质组水平上以低表达相关性调节,主要在洪水后期起作用。参与活性氧(ROS)清除的基因,线粒体代谢,发育也完全在转录组水平上受到调节,但它们的表达水平在洪水后恢复时高度上调。此外,与氧化还原相关的基因/蛋白质的综合表达谱,荷尔蒙,和转录因子也进行了研究。最后,讨论了M.laxiflora应对洪水和洪水后恢复的监管网络。这些发现加深了我们对洪水胁迫的分子机制的理解,并阐明了在洪水区保存M.laxiflora和其他濒危植物的基因和途径。
    Flooding is one of the major environmental stresses that severely influence plant survival and development. However, the regulatory mechanisms underlying flooding stress remain largely unknown in Myricaria laxiflora, an endangered plant mainly distributed in the flood zone of the Yangtze River, China. In this work, transcriptome and proteome were performed in parallel in roots of M. laxiflora during nine time-points under the flooding and post-flooding recovery treatments. Overall, highly dynamic and stage-specific expression profiles of genes/proteins were observed during flooding and post-flooding recovery treatment. Genes related to auxin, cell wall, calcium signaling, and MAP kinase signaling were greatly down-regulated exclusively at the transcriptomic level during the early stages of flooding. Glycolysis and major CHO metabolism genes, which were regulated at the transcriptomic and/or proteomic levels with low expression correlations, mainly functioned during the late stages of flooding. Genes involved in reactive oxygen species (ROS) scavenging, mitochondrial metabolism, and development were also regulated exclusively at the transcriptomic level, but their expression levels were highly up-regulated upon post-flooding recovery. Moreover, the comprehensive expression profiles of genes/proteins related to redox, hormones, and transcriptional factors were also investigated. Finally, the regulatory networks of M. laxiflora in response to flooding and post-flooding recovery were discussed. The findings deepen our understanding of the molecular mechanisms of flooding stress and shed light on the genes and pathways for the preservation of M. laxiflora and other endangered plants in the flood zone.
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  • 文章类型: Journal Article
    Whitmania pigra has been used as a traditional Chinese medicine (TCM) for promoting blood circulation, alleviating blood coagulation, activating meridians and relieving stasis for several hundred years. However, the therapeutic components of this species, especially proteins and peptides were poorly exploited. Until now only a few of them were obtained by using chromatographic isolation and purification. In recent decade, transcriptome techniques were rapidly developed, and have been used to fully reveal the functional components of many animal venoms. In the present study, the cDNA of the salivary gland of Whitmania pigra was sequenced by illumina and the transcriptome was assembled by using Trinity. The proteome were analysed by LC-MS/MS. Based on the data of the transcriptome and the proteome, a potential antiplatelet protein named pigrin was found. Pigrin was cloned and expressed using P. pastoris GS115. The antiplatelet andantithrombotic bioactivities of pigrin were tested by using aggregometer and the rat arterio-venous shunt thrombosis model, respectively. Thebleeding time of pigrin was measured by a mice tail cutting method. The docking of pigrin and protease-activated receptor 1 (PAR1) or collagen were conducted using the ZDOCK Server. Pigrin was able to selectively inhibit platelet aggregation stimulated by PAR1 agonist and collagen. Pigrin attenuated thrombotic formation in vivo in rat, while did not prolong bleeding time at its effective dosage. There are significant differences in the key residues participating in binding of Pigrin-Collagen complex from Pigrin-PAR1 complex. In conclusion,a novel PAR1 inhibitor pigrin was found from the leech Whitmania pigra. This study helped to elucidate the mechanism of the leech for the treatment of cardiovascular disorder.
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