BRC-BIO: biochemical activation mechanisms of arabidopsis GCN2 and translational control by reactive oxygen

This National Science Foundation (NSF) Biological Sciences program (CFDA 47.074) Project Grant award provides $497,039 to Valdosta State University to investigate the biochemical mechanisms by which reactive oxygen species activate the GCN2 protein kinase and its role in regulating protein synthesis in Arabidopsis thaliana as a model plant system. The research aims to enhance understanding of plant stress responses and translational control mechanisms, which could contribute to developing strategies for improving crop resilience. In addition to the scientific research, the project includes initiatives to promote diversity and inclusion in STEM education, provide training opportunities for undergraduate and graduate students, and engage local high school students and educators in hands-on STEM activities. The award was co-funded by the NSF's Division of Molecular and Cellular Biosciences and Division of Biological Infrastructure, and the project period runs from June 15, 2024 to May 31, 2027.

Abstract

This research project delves into the complex realm of stress responses of plants, which is pivotal for improving crop resilience and ensuring a stable food supply. Imagine plants as resilient heroes that encounter and overcome challenges associated with a dynamic landscape that may include changes in temperature and light levels and the presence of herbivores. A highly reactive class of compounds, reactive oxygen species (ROS), serves as alert signals, triggering plants to activate defense mechanisms. By deciphering how plants interpret and react to these stress signals at the level of protein synthesis, we can potentially develop better strategies for enhancing crop resilience to environmental pressures, ultimately contributing to sustainable agriculture. Beyond its scientific significance, this research endeavor creates opportunities for undergraduate and graduate students from diverse backgrounds to engage in scientific exploration. Through tailored educational programs and hands-on workshops for high school students and educators, the project not only sparks curiosity but also highlights the captivating intricacies of plant stress responses. By igniting passion for science and promoting inclusivity, the project aims to inspire future generations of scientists while fostering diversity in science, technology, engineering, and mathematics (stem) fields. This project investigates the mechanisms by which reactive oxygen species (ROS) activate the highly conserved protein kinase, general control of nonderepressible 2 (GCN2), in arabidopsis thaliana. GCN2 plays a crucial role in regulating protein synthesis (translational control), a fundamental process in all organisms, by phosphorylating the translation initiation factor, EIF2ALPHA. Despite the well-documented involvement of GCN2 in a wide variety of stress responses, the precise mechanisms of its activation in plants remain elusive. Building upon our previous findings that link ROS accumulation to rapid GCN2 activation, this study aims to elucidate the biochemical pathways mediating ROS-GCN2 signaling. The project involves in vitro and EX vivo kinase assays coupled with advanced proteomics techniques to dissect the mode of ROS-mediated GCN2 activation. Furthermore, genome-wide analyses of mrna-ribosome loading will be employed to discern the specific impact of GCN2-EIF2ALPHA signaling on translation control under ROS stress. These investigations will enhance our understanding of plant stress responses and contribute to broader knowledge in translational control mechanisms in plants, as well as across all eukaryotes. Additionally, the project's broader impacts include initiatives to promote diversity and inclusion in stem education, providing valuable training opportunities for undergraduate and graduate students and engaging local high school educators and their students in hands-on stem activities. This award was CO-funded by the division of molecular and cellular biosciences and the division of biological infrastructure. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the foundation's intellectual merit and broader impacts review criteria.- subawards are not planned for this award.