Rediscovering hypoxia though RNA regulation

缺氧是活细胞和生物体常见的应激状态，与癌症等疾病有关。在缺氧期间，细胞通过激活一系列适应性反应来补偿氧气的缺乏，以满足代谢、生物能量和氧化还原的需求。这些反应是多方面的，需要精确调节基因表达，并涉及几个细胞途径。转录机制如未折叠蛋白反应（UPR）、哺乳动物雷帕霉素靶蛋白（mTOR）信号传导和缺氧诱导因子（HIF）的基因调控已被广泛研究。然而，缺氧时基因的表达也受到转录后的高度影响，RNA结合蛋白（RBP）在控制这些过程（如mRNA的稳定性和翻译效率）中起着关键作用.此外，最近的数据提出了一个有趣的新想法，即酶也可以作为RNA结合蛋白发挥作用，RNA结合可能以变构方式调节它们的活性;然而，关于其机制的细节，或者是否在缺氧时会发生这种情况，我们知之甚少。该项目的目标是：目标1。通过无偏的蛋白质组学方法和定量质谱分析，揭示RBP在缺氧条件下的RNA结合活性变化。(Year 2 -注：第1年为轮换项目）。目标二。通过代谢组学方法确定代谢酶的RNA结合变化与其代谢活性之间的联系。(Year 2）。目标3：目标1和2中有关RBP和缺氧代谢的信息将揭示机制分析应重点关注哪些RPB候选物，包括典型的RBP和缺氧期间结合RNA的酶。这些将被探索使用细胞，生物化学，生物物理和结构的方法来理解RNA调控的精确机制。（3-4年级）。为此，我们将采用结构、生物物理和细胞生物学相结合的方法，结合最先进的多组学技术，

Hypoxia is a common stress condition for living cells and organisms, and is linked tomany diseases such as cancer. During hypoxia, cells compensate for the lack ofoxygen, by activating a series of adaptive responses to satisfy the metabolic,bioenergetic and redox demands. These responses are multifaceted requiring preciseregulation of gene expression and involving several cellular pathways. Transcriptionalmechanisms like unfolded protein response (UPR), mammalian target of rapamycin(mTOR) signalling, and gene regulation by hypoxia inducible factor (HIF) have beenextensively studied. However gene expression during hypoxia is also highly affectedpost-transcriptionally and RNA binding proteins (RBPs) are critical in controlling theseprocesses (e.g. mRNA stability and translation efficiency). Moreover recent data haveproposed the intriguing and novel idea that enzymes can also function as RNA bindingproteins, and that RNA binding would modulate their activity probably in an allostericfashion; however very little is known about the mechanistic details, or whether thishappens during hypoxia. The aims of this project are: Aim 1. To reveal changes in RNAbinding activity of RBPs in response to hypoxia through an unbiased proteomicsapproach with quantitative mass spectrometry analysis. (Year 2 - Note Year 1 is forrotation projects). Aim 2. To define the link between changes in RNA binding ofmetabolic enzymes and their metabolic activity through metabolomics approaches.(Year 2). Aim 3. The information on RBPs and hypoxia metabolism from aim 1 and 2will reveal which RPB candidates to focus on for mechanistic analysis, includingcanonical RBPs and enzymes that bind RNA during hypoxia. These will be exploredusing cellular, biochemical, biophysical and structural approaches to understand theprecise mechanisms of RNA regulation.( Year 3-4). For this we will employ a combinedstructural, biophysical, and cellular biology approach, in conjugation with state-of-artmulti-omics