Generation of cell lines with tetracycline-regulated autophagy and a role for autophagy in controlling cell size
Generation of cell lines with tetracycline-regulated autophagy and a role for autophagy in controlling cell size
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DOI：
10.1016/j.febslet.2006.04.008
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发表时间：
2007-06-19
期刊：
影响因子：
3.5
通讯作者：
Mizushima, Noboru
中科院分区：
文献类型：
--
作者：
Hosokawa, Nao;Hara, Yukichi;Mizushima, Noboru
自噬是细胞内的整体降解系统。我们建立了小鼠成纤维细胞系，将Tet-off系统与Atg 5（-1-）小鼠胚胎成纤维细胞系偶联，以人工调节自噬能力。在多西环素（Dox）的存在下，Atg 5的表达被完全抑制，这些细胞是自噬缺陷的。去Dox后，自噬能力在6 h内恢复。非常低水平的Atg 5可以诱导自噬活性状态。我们应用这个新系统来研究自噬对控制细胞大小的贡献。在不能进行自噬的细胞中，响应于饥饿的细胞尺寸减小被显著抑制。所产生的细胞系将是有用的试剂，为未来的机制研究的调节和生理意义的自噬。(c)2006年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
Autophagy is an intracellular bulk degradation system. We established mouse fibroblast lines coupling the Tet-off system with an Atg5(-1-) mouse embryonic fibroblast line to artificially regulate autophagic ability. In the presence of doxycycline (Dox), Atg5 expression was completely suppressed and these cells were autophagy-defective. After removal of Dox, autophagic ability was restored within 6 h. Very low levels of Atg5 could induce an autophagy competent state. We applied this novel system to examine the contribution of autophagy to controlling cell size. Cell size reduction in response to starvation was significantly inhibited in cells unable to undergo autophagy. The generated cell lines will be useful reagents for future mechanistic studies into the regulation and physiologic significance of autophagy. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.