Millen Jonathan I, Krick Roswitha, Prick Tanja, Thumm Michael, Goldfarb David S
Department of Biology, University of Rochester, Rochester, NY 14627, USA.
Autophagy. 2009 Jan;5(1):75-81. doi: 10.4161/auto.5.1.7181.
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.
细胞核的逐片微自噬（PMN）选择性地清除并降解酿酒酵母细胞核的小片段。被称为核-液泡（NV）连接的细胞器间接触位点通过为PMN小泡和囊泡的生物发生建立一个平台来决定PMN的选择性。使用绿色荧光蛋白标记的报告基因，可通过荧光显微镜观察PMN结构；然而，这种方法最好辅以PMN特异性货物降解的定量免疫印迹分析。总之，这些分析应有助于在不同遗传背景、生理状态和环境条件下进一步研究这个迷人但了解甚少的自噬过程。
