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Published on: July 18, 2011
Measuring piecemeal microautophagy of the nucleus in Saccharomyces cerevisiae
Jonathan I Millen1, Roswitha Krick, Tanja Prick
1Department of Biology, University of Rochester, Rochester, NY 14627, USA.
Abstract:
Piecemeal microautophagy of the nucleus (PMN) selectively removes and degrades small fragments of the Saccharomyces cerevisiae nucleus. Inter-organelle contact sites called nucleus-vacuole (NV) junctions determine the selectivity of PMN by establishing a platform for the biogenesis of PMN blebs and vesicles. PMN structures can be observed by fluorescence microscopy using GFP-tagged reporters; however, this approach is best supported with quantitative immunoblot assays of PMN-specific cargo degradation. Together, these assays should facilitate the further study of this fascinating but poorly understood autophagic process in different genetic backgrounds, physiological states, and environmental conditions.
Insights
Piecemeal microautophagy of the nucleus (PMN) selectively degrades nuclear fragments in yeast. Nucleus-vacuole junctions are key to this process, enabling further study of this autophagic pathway.
Area of Science:
- Cell Biology
- Molecular Biology
- Autophagy Research
Background:
- Piecemeal microautophagy of the nucleus (PMN) is a selective autophagic process in Saccharomyces cerevisiae.
- PMN targets and degrades small nuclear fragments.
- Nucleus-vacuole (NV) junctions are implicated in the selectivity of PMN.
Purpose of the Study:
- To elucidate the role of nucleus-vacuole junctions in piecemeal microautophagy of the nucleus.
- To establish robust methods for studying PMN in yeast.
- To facilitate future research into PMN under diverse conditions.
Main Methods:
- Utilizing fluorescence microscopy with GFP-tagged reporters to visualize PMN structures.
- Employing quantitative immunoblot assays to measure PMN-specific cargo degradation.
- Investigating PMN in various genetic backgrounds, physiological states, and environmental conditions.
Main Results:
- Nucleus-vacuole (NV) junctions act as platforms for PMN bleb and vesicle biogenesis.
- GFP-tagged reporters allow visualization of PMN.
- Quantitative immunoblots provide a method for assessing PMN cargo degradation.
Conclusions:
- NV junctions are critical for the selectivity of piecemeal microautophagy of the nucleus.
- A combination of fluorescence microscopy and immunoblot assays supports the study of PMN.
- These methods will enable comprehensive investigation of PMN in yeast.
