The spatial organization of lipid synthesis in yeast: Large-scale
GFP-tagging and high-resolution microscopy.
Klaus Natter (1),
Iskandar Dib (1), Alexander Faschinger (1), Anita Jandrositz (1), Steven
McCraith (2), Stanley Fields (2), Sepp D. Kohlwein (1)
(1)
IMBM-Biochemistry, University of Graz, Schubertstr. 1, Graz, A8010,
Austria; (2) Howard Hughes Medical Institute, University of Washington,
Seattle
Yeast is an attractive model organism for molecular
biological research, but represents a particular challenge to
microscopists due to its small overall dimensions of 5-8 µm. However,
knowledge about the localization of proteins in a cell provides
important information as to their function. We make use of a genome-wide
approach for protein localization studies in yeast, based on GFP-tagging
by recombination cloning, and high resolution confocal microscopy. A
central focus of our studies is the identification of the spatial
organization of proteins involved in lipid metabolism. GFP-tagged
proteins associated with lipid droplets and presumably involved in fatty
acid and neutral lipid metabolism are characterized as to their
subcellular distribution and dynamics upon conditions of lipid
mobilization. From the microscopic large-scale analysis of GFP-tagged
proteins in vivo, novel factors of unknown function are selected based
on their localization to lipid droplets, and are further characterized
with respect to their predicted participation in lipid metabolism.
Microscopic images, together with experimental setup and documentation,
are stored in a web accessible Oracle-based Yeast Protein Localization
Database, YPL.db.
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