An Integrative Approach to Elucidate Systemic RNA Trafficking

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How basic processes in individual cells within a multicellular organism are integrated to enable development, function and response to the environment is a central issue in modern biology. Cell-to-cell trafficking of RNA is a recently discovered mechanism that controls gene regulation, viral infection, cellular antiviral responses, and cell-to-cell communication. This project investigates how an RNA produced in a cell is selected for transport into neighboring cells and even to distant organs in a plant, a basic mechanistic question of fundamental importance to understand trafficking regulation for all RNAs. Potato spindle tuber viroid (PSTVd) infection will be used as a simple experimental model to test the hypothesis that an RNA has distinct structural motifs to interact with cellular factors to potentiate trafficking across successive cellular boundaries from the plant surface to the centrally located vascular tissue for long-distance transport. Specifically, the project will (1) analyze a series of trafficking-defective mutants of PSTVd to identify the RNA motifs mediating trafficking through different cellular boundaries, (2) test the predicted three-dimensional structural model of a trafficking motif through extensive mutational analyses, and (3) determine the role of a plant protein in PSTVd trafficking. Results from the project will provide groundbreaking insights about how an infectious RNA has evolved distinct structural elements to use cellular machinery to establish infection, and help develop a conceptual framework to study the mechanisms of RNA trafficking underlying many biological processes. This project will include training of undergraduate and graduate students. This training will expose students to cutting-edge cross-disciplinary research approaches to study fundamental biological problems. The project will also enhance science education for elementary/middle/high school students and secondary school teachers, including traditionally underrepresented groups.