NSF/MCB-BSF: RNA quality control in the chloroplast

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One of the main avenues to optimizing plant performance is to improve their ability to harvest the sun’s energy. Plant cells contain chloroplasts where light energy is harvested and converted into sugars and ultimately into food, feed and fiber. The chloroplast contains its own DNA and genes that must function correctly for photosynthesis to occur. Like all biological processes, these functions are prone to errors that must be corrected for plants to function. This project seeks to understand the fundamental yet mysterious process by which this quality control is accomplished. Quality control of this type is not specific to chloroplasts, nor to plants, and therefore advances made in this project will be applicable to other living systems. The project also creates opportunities for training diverse undergraduates in experimental and computational techniques, including plant genetic engineering and bioinformatic analysis. Students will also be taught communications skills and will be exposed to a variety of career options. This training will occur through mentoring and teamwork, which are important transferable skills for early career scientists. In chloroplasts, RNA quality control is of paramount importance because transcription itself is relatively unregulated. This project builds on biochemical, genetic and computational findings in Arabidopsis, focusing on the essential RNA quality control enzyme, RNase J, which has been shown to trim 5’ and 3’ RNA termini and to eliminate deleterious antisense RNA. This project seeks to discover how RNase J is regulated, and how its specificity is imparted. The two major approaches are first, to create transgenic Arabidopsis lines that express versions of RNase J with informative site-directed mutations or deletions, and second, to biochemically measure the activities of RNaseJ in vitro. Overall, the experiments should advance knowledge of how the chloroplast transcriptome is shaped, and provide insights into mechanisms that discriminate transcripts among large populations of RNA molecules.This award was co-funded by the Genetic Mechanisms program of the Division of Molecular and Cellular Biosciences, and the Plant Genome Research Program of the Division of Integrative Organismal Systems in the Biological Sciences Directorate.This collaborative US/Israel project is supported by the US National Science Foundation and the Israeli Binational Science Foundation.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.