Rediscovering hypoxia though RNA regulation

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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