Reference: Devenish RJ, et al. (2008) Monitoring organelle turnover in yeast using fluorescent protein tags. Methods Enzymol 451:109-31

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Abstract


This chapter describes the use of fluorescent protein (FP) tags to monitor autophagic degradation of various organelles in the yeast S. cerevisiae. The advantages of specific targeting of FPs to different organelles are the ability to follow, under specific conditions: (1) the localization and/or distribution of the labeled (fluorescent) target organelle in relation to the vacuole, and (2) autophagic degradation of the target organelle in a single live cell. Currently, our mechanistic knowledge of organellophagy as a selective form of autophagy is still quite limited. Accordingly, continued efforts are needed to elucidate the detailed mechanism and to uncover the basis of selectivity of organellophagy. FPs have long been recognized as powerful tools for uncovering dynamic processes in eukaryotic cells. They serve as excellent markers of organelles for studies of autophagic degradation and, as such, have the potential to greatly expand our knowledge of organellophagy in various model systems (from yeast to higher eukaryotes). The use of organellespecific FPs as a means to follow autophagic degradation of different organelles and its dependency on the characterized autophagic machinery is, relatively speaking, still in its infancy. What is presented here is not a formulaic set of rules, but guidelines for the use of FP technology in the context of autophagy. Further developments and specific uses of different FPs to ask new questions are sure to be forthcoming.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Devenish RJ, Prescott M, Turcic K, Mijaljica D
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