Measuring piecemeal microautophagy of the nucleus in Saccharomyces cerevisiae.
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.
Millen
Jonathan I
JI
Department of Biology, University of Rochester, Rochester, NY 14627, USA.
Krick
Roswitha
R
Prick
Tanja
T
Thumm
Michael
M
Goldfarb
David S
DS
D001343:Autophagy; D001426:Bacterial Proteins / Q000378:metabolism; D021961:Cell Membrane Structures / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D003584:Cytological Techniques / Q000379:methods*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015151:Immunoblotting; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
D001426:Bacterial Proteins; D008164:Luminescent Proteins; C499847:Nvj1 protein, S cerevisiae; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins