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学科主题: 基础医学
题名:
High-throughput functional screening for autophagy-related genes and identification of TM9SF1 as an autophagosome-inducing gene
作者: He, Pengfei; Peng, Zhi4; Luo, Ye; Wang, Lan2,3; Yu, Peng; Deng, Weiwei; An, Yunqing4; Shi, Taiping1,2,3; Ma, Dalong2,3
关键词: high-throughput screening ; autophagy ; LC3 ; TM9SF1 ; automated fluorescence microscopy system
刊名: AUTOPHAGY
发表日期: 2009
卷: 5, 期:1, 页:52-60
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Cell Biology
研究领域[WOS]: Cell Biology
关键词[WOS]: MONITORING AUTOPHAGY ; CELL-DEATH ; PROTEIN ; MARKER ; MONODANSYLCADAVERINE ; APOPTOSIS ; HOMOLOG ; MODEL ; YEAST ; LC3
英文摘要:

Autophagy, a tightly regulated process responsible for the bulk degradation of most long-lived proteins and some organelles, is associated with several forms of human diseases including cancer, neurodegenerative disease and cardiomyopathies. However, the molecular machinery involved in autophagy in mammalian cells remains poorly understood. Here, we describe a high-throughput, cell-based functional screening platform, based on an automated fluorescence microscopy system, which enables acquiring and quantitatively analyzing images of GFP-LC3 dots in cotransfected cells. From a library of 1,050 human cDNA clones, we identified three genes (TM9SF1, TMEM166 and TMEM74) whose overexpression induced high levels of autophagosome formation. In particular, overexpression of TM9SF1, which colocalized with LC3 according to the confocal assay, led to a significant increase in the number of GFP-LC3 dots. The results of transmission electron microscopy and immunoblotting to examine LC3-II levels further confirmed the ability of TM9SF1 to induce autophagy. Furthermore, knockdown of TM9SF1 expression by RNA interference could hamper starvation-induced autophagy. The functional screening platform therefore can be applied to high-throughput genomic screening candidate autophagy-related genes, which would provide new insights into underlying molecular mechanisms that may regulate autophagy in mammalian cells.

语种: 英语
所属项目编号: 2006AA02A305
项目资助者: National High Technology Research and Development Program of China
WOS记录号: WOS:000262844800006
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/65327
Appears in Collections:基础医学院_免疫学系_期刊论文

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作者单位: 1.Capital Med Univ, Dept Immunol, Beijing, Peoples R China
2.Chinese Natl Human Genome Ctr, Beijing Funct Genom Lab 3, Beijing 100176, Peoples R China
3.Peking Univ, Hlth Sci Ctr, Sch Basic Med Sci, Lab Med Immunol, Beijing 100871, Peoples R China
4.Peking Univ, Ctr Human Dis Genom, Beijing 100871, Peoples R China

Recommended Citation:
He, Pengfei,Peng, Zhi,Luo, Ye,et al. High-throughput functional screening for autophagy-related genes and identification of TM9SF1 as an autophagosome-inducing gene[J]. AUTOPHAGY,2009,5(1):52-60.
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