Three-dimensional analysis of protein aggregate body in Saccharomyces cerevisiae cells.
Modified genes of peroxisomal isocitrate lyase of Candida tropicalis ( CT- ICL) were constructed and expressed in Saccharomyces cerevisiae cells. We observed subcellular localization of expressed products of the mutant CT- ICL genes by immunoelectron microscopy. An unknown structure termed a protein aggregate body (PAB) storing the expressed product was observed in cytoplasm in various mutants (Kamasawa et al. (1996) J. Electron Microsc. 45: 491-497). We chose two typical cells harbouring the mutant ICL genes delta 550 and delta 237-339 to analyse the ultrastructure and three-dimensional (3D) structure of PABs. The PABs had a homogeneous matrix with a wavy periphery in the cell image using a high-pressure freezing fixation method. Although PABs could not be separated from the cytoplasm or mitochondria under a confocal fluorescence microscope, 3D reconstruction of serial electron micrographs clearly showed the PAB was an independent structure of varying size and had the shape of an incomplete sphere. A cell was sometimes observed to have multiple PABs.[1]References
- Three-dimensional analysis of protein aggregate body in Saccharomyces cerevisiae cells. Kamasawa, N., Yoshida, T., Ueda, M., Tanaka, A., Osumi, M. Journal of electron microscopy. (1999) [Pubmed]
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