Equipment Supplement for R35 GM144059 "Writers and Erasers of Ubiquitin Moieties in Control of Cell-to-Cell Transport in Plants"

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PROJECT SUMMARY Intercellular transport is central to the life cycle of multicellular organisms, and its molecular mechanisms and their regulation have been extensively studied in different model systems, including plants and animals. Yet, one key aspect of cell-to-cell transport regulation apparently escaped attention: the potential involvement of ubiquitylation/deubiquitylation (and the corresponding ubiquitin ligases and deubiquitinases). In our parental project, we have obtained evidence for the existence of such a regulatory system and are characterizing its molecular components and mechanism(s) of action. This work heavily depends on the use of a confocal laser scanning microscope to detect and follow the fate of fluorescently tagged proteins in plant tissues. Based on the current and projected trajectory of the project, there is an urgent need for improvement of our confocal imaging quality, i.e., resolution and signal-to-noise ratio, to enhance the quality of acquired data and their interpretation. To address this critical technical aspect of the project, we propose acquiring a Zeiss Airyscan 2 detection system with a Zeiss 60A R2 offline analysis workstation that can be incorporated into our existing Zeiss LSM 900 machine and provide significant improvement to its imaging and image analysis capabilities.