A novel eukaryotic RNA thermoswitch: molecular function and biotech applications

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Responsiveness to environmental stresses, including temperature, is a crucial feature for sessile organisms such as plants so they can adjust their growth and development accordingly. We have recently discovered a novel plant RNA ThermoSwitch (pRTS) that is responsible for day-time rhythmic growth of the model flowering plant Arabidopsis (Chung* et al, May 2020, Nature Plants, with accompanying News and Views highlight). In contrast to more established protein-based thermo-sensors, the pRTS is an RNA element, which directly regulates protein synthesis through rapid conformation changes in response to temperature fluctuations. Through a combination of computation and experimental assays, we have demonstrated that the pRTS drives the protein synthesis of several transcription factors such PIF7, WRKY22 and HSFA2, that control the expression of many proteins, within minutes after the temperature changes from 17C to 27C. Further investigation on the role of PIF7 revealed that it drives transcription for several growth regulators, resulting in day-time growth of Arabidopsis.However, this previous work just scratched the surface of understanding the biological function of pRTS. Many very important questions remained unanswered, especially as to how pRTSs manipulate translation dynamics to achieved enhanced protein synthesis within such a short timeframe. Further, the newly discovered pRTSs have great biotechnological potential as energy-efficient rapid-inducers for heterologous protein expression for molecular pharming - an appealing system for high throughput production in response to immediate needs such as vaccine production during outbreaks and pandemics. We will address these two questions through the following objectives: 1. We will decipher pRTS-mediated translation dynamics. This information will further our understanding of inherent differences between the plant and animal translation machinery and will provide a molecular explanation for the more thermo-responsive nature of plants. 1a. We will determine whether pRTSs are phylogenetically conserved within the plant kingdom.1b. We will dissect the molecular dynamics of pRTS-dependent translation by mapping all translation complexes (scanning, initiating, elongating and terminating complexes) before and after pRTS activation.2. We will develop an energy-efficient thermo-inducible high-level expression system for molecular pharming.2a. We will identify optimal features of pRTSs for rapid thermo-induced protein synthesis2b. We will develop a thermo-inducible high-yield heterologous gene expression system that can be utilised by the biopharma industry2c. Proof of concept strategy: To confirm efficiency, yield and efficacy in planta with the system developed in 2b.This research is exciting for several reasons. The discovery of plant ThermoSwitches opens up a whole new avenue for adaptive response to temperature changes in plants at the level of protein synthesis. We are ideally poised to exploit this new research direction. Understanding the translation dynamics driven by the plant ThermoSwitch may also help explain why translation machinery in the animal kingdom is less responsive to temperature change. Importantly, biotechnological utilisation of pRTSs could provide an ideal system for rapid vaccine production in an economical manner when demand is urgent.