RNA turnover in plant stress responses

植物逆境反应中的RNA周转

• 批准号: 433113158
• 负责人: Professor Dr. Sascha Laubinger
• 金额: --
• 依托单位: Abteilung Pflanzengenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/433113158?language=en
• 关键词: RNA; turnover; plant; stress; responses

Modulating the stability of RNAs is a key feature of gene expression regulation. While housekeeping genes usually produce highly stable RNAs, stress-responsive genes often encode RNAs with low stability. This allows rapid RNA turnover; a feature particularly important for plants, which have to adapt quickly to changing environmental conditions and stresses. The pathways that execute RNA degradation in plants are well understood. However, it is largely unknown how stresses destabilize specific RNAs, and how target specificity of RNA degrading enzymes is controlled.To address these questions, we will apply our recently developed ERIC-sequencing technology to stressed and unstressed Arabidopsis thaliana plants. Traditionally, RNA decay over time and RNA half-lives have been determined after harsh treatments with highly toxic transcriptional inhibitors such as actinomycin D and cordycepin. We developed a novel method (ERIC-seq), which employs metabolic RNA labeling using the non-toxic 5-ethynyl uridine (5-EU) combined with RNA sequencing. This allowed us for the first time to determine RNA stability non-invasively in plants. We will apply this technique to study the stability of nuclear and chloroplast-encoded RNAs under a diverse set of abiotic stress conditions. In addition, we will study the role of specific cytosolic and plastidic RNA degradation and stabilization pathways under selected stress conditions. This approach enables us to identify stress-specific RNA sequence features, that (1) ensure RNA stabilization and destabilization under abiotic stress conditions, and that (2) determine the specificity of RNA degradation pathways for subpopulations of RNAs. Such sequence elements will be functionally studied, linked to specific endonucleases, and used to identify novel trans-regulatory RNA stability factors. As a major deliverable, our approach will identify the impact of RNA turn-over on gene regulation during stress on a transcriptome-wide scale.

调节RNA的稳定性是基因表达调控的关键特征。虽然管家基因通常产生高度稳定的RNA，但应激反应基因通常编码稳定性低的RNA。这使得RNA能够快速周转;这一特征对于植物特别重要，因为植物必须快速适应不断变化的环境条件和压力。在植物中执行RNA降解的途径是众所周知的。然而，这在很大程度上是未知的压力是如何不稳定的特定RNA，以及如何控制RNA降解酶的靶特异性。为了解决这些问题，我们将应用我们最近开发的ERIC测序技术，强调和非强调拟南芥植物。传统上，RNA随时间的衰减和RNA半衰期已经在用高毒性转录抑制剂如放线菌素D和虫草素进行苛刻处理后确定。我们开发了一种新的方法（ERIC-seq），该方法使用无毒的5-乙炔基尿苷（5-EU）结合RNA测序进行代谢RNA标记。这使我们首次能够非侵入性地确定植物中RNA的稳定性。我们将应用这种技术来研究核和叶绿体编码的RNA在不同的非生物胁迫条件下的稳定性。此外，我们将研究特定的胞质和质体RNA降解和稳定途径在选定的压力条件下的作用。这种方法使我们能够识别胁迫特异性RNA序列特征，（1）确保RNA在非生物胁迫条件下的稳定和不稳定，以及（2）确定RNA亚群的RNA降解途径的特异性。将对这些序列元件进行功能研究，将其与特定的核酸内切酶连接，并用于鉴定新型反式调节RNA稳定因子。作为一个主要的可交付成果，我们的方法将确定RNA周转对基因调控的影响，在压力下在转录组范围内。